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Re: [DIYbio] Re: THC legislation is next week

Would you say Marijuana is more dangerous than chemotherapy? Seems immoral and illogical.

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Re: [DIYbio] Re: THC legislation is next week

On 10/31/2012 05:04 PM, Nathan McCorkle wrote:
> We commonly discuss open source laws here, gene
> patents, microcontrollers. I think laws about gene patenting or laws
> about gene outlawing (which this pretty much is) is relevant here.

Yep.

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Re: [DIYbio] Re: THC legislation is next week

On Wed, Oct 31, 2012 at 2:48 PM, Josiah Zayner <josiah.zayner@gmail.com> wrote:
> Taxol has proven way more effective against cancer and can be grown in your
> backyard. http://en.wikipedia.org/wiki/Paclitaxel

The problem with anti-cancer claims is that cancer is a broad term for
many specific cellular malfunctions. We need to study all the options
for treatment.

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Re: [DIYbio] I am a programmer looking for webapp idea that can be done in 5 weeks!

on linux (and I think windows VS), openCV had its own implementation
for displaying pics and sliders, so you shouldn't need to use any
VS/mono GUI stuff.

On Wed, Oct 31, 2012 at 2:25 PM, Sebastian S. Cocioba
<scocioba@gmail.com> wrote:
> All this gtk# sillyness for emguCV in monodevelop is gonna take me some time to get used to. Would you mind if I do it in C# visual 2012 and flush out all the concepts and needs of the app and then port it over to mono? The windows monodevelop pack comes with a migration wizard that converts vs to mono. The pInvoke thing is still an issue in mono but there are work arounds. Just a thought.
>
> Sebastian S Cocioba
> CEO & Founder
> New York Botanics, LLC
>
> Sent via Mobile E-Mail
>
> On Oct 31, 2012, at 1:23 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
>
>> Sebastian, if you want to do some image processing in .NET or Mono
>> using openCV (I've done it before in Visual Studio using an online
>> tutorial, it's a PITA more than on linux but it works) I can then move
>> the openCV code to C/C++ and throw it under a django or maybe flask
>> web GUI (i've been meaning to play with flask).
>>
>>
>> On Mon, Oct 29, 2012 at 12:56 PM, Sebastian S. Cocioba
>> <scocioba@gmail.com> wrote:
>>> I solely do .NET programming so my offer was for any windows users. The
>>> reason why its easy is because i already have code for doing object recog
>>> with things broken out so a few tweaks later and it fits. Only catch is that
>>> it will be a visual studio project and whatnot. I do want to switch over to
>>> more cross platform methods like direct openCV use in C++. Sry for any
>>> confusion.
>>>
>>>
>>> Sebastian S Cocioba
>>> CEO & Founder
>>> New York Botanics, LLC
>>>
>>> Sent via Mobile E-Mail
>>>
>>> On Oct 29, 2012, at 3:49 PM, Bryan Bishop <kanzure@gmail.com> wrote:
>>>
>>> On Mon, Oct 29, 2012 at 2:45 PM, Sebastian S. Cocioba <scocioba@gmail.com>
>>> wrote:
>>>>
>>>> Hmm if anyone would be interested in something like that in a windows app
>>>> I could whip it up quickly
>>>
>>>
>>> I am confused.. why is it easier for you to make something that's *not*
>>> cross-platform? What?
>>>
>>> - Bryan
>>> http://heybryan.org/
>>> 1 512 203 0507
>>>
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>>
>>
>> --
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>>
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Re: [DIYbio] Re: THC legislation is next week

As was mentioned, the legal status has large implications for
scientific study. We commonly discuss open source laws here, gene
patents, microcontrollers. I think laws about gene patenting or laws
about gene outlawing (which this pretty much is) is relevant here. If
the plant itself is really the problem, let's start a thread about
reproducing some of the THC tobacco hairy root culture work that's
been done. I'm pretty sure we're talking about a legitimate scientific
roadblock here.

Oil is much better at making textiles than hemp is (at least in terms
of strength I think, and probably mass in to clothes out), maybe if we
don't have a decent biofuel/bio-oil replacement soon we'll see an
upturn in hemp fiber utility, but I am not counting on that.

Maybe the seeds just contain omega-3s?

On Wed, Oct 31, 2012 at 2:48 PM, Josiah Zayner <josiah.zayner@gmail.com> wrote:
> Taxol has proven way more effective against cancer and can be grown in your
> backyard. http://en.wikipedia.org/wiki/Paclitaxel
> You can make textiles from Grass.
> Omega-3 fatty acids are more effective at reducing cholesterol.
> Salicylic acid aka aspirin is a plant horomone and gives relief to
> individuals suffering from a number of conditions.
>
> Really, believe the lies fueled by racial profiling? LOL
>
> I never said anything for or against legalization of marijuana. All I said
> was that a discussion about the legalization of marijuana has no place in
> DIY Bio forum. No one is discussing the scientific efficacy of marijuana and
> no one plans on using it in any scientific way. I _personally_ just think it
> is sad that the number one topic of the day has nothing to do with science
> or DIY Bio but drug legalization.
>
>
> On Wednesday, October 31, 2012 4:25:35 PM UTC-5, GeneralOya wrote:
>>
>> Hmmm not on topic of DIYBio huh. Well it's a proven effective cancer
>> medicine, which folks can grow in their home/yard. What more do you want?
>>
>> You can also make fuel from it.
>> You can also make textiles from it.
>> The seeds are effective in reducing cholesterol.
>> It gives immediate relief to individuals suffering from a number of
>> conditions.
>>
>> So believe the lies from an era of prohibition fueled racial profiling, or
>> do the research. It's a fabulous plant. I'm glad to see a few folks chiming
>> in regardless of where they stand.
>>
>> Ryan
>
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Re: [DIYbio] Re: THC legislation is next week

Taxol has proven way more effective against cancer and can be grown in your backyard. http://en.wikipedia.org/wiki/Paclitaxel
You can make textiles from Grass.
Omega-3 fatty acids are more effective at reducing cholesterol.
Salicylic acid aka aspirin is a plant horomone and gives relief to individuals suffering from a number of conditions.

Really, believe the lies fueled by racial profiling? LOL

I never said anything for or against legalization of marijuana. All I said was that a discussion about the legalization of marijuana has no place in DIY Bio forum. No one is discussing the scientific efficacy of marijuana and no one plans on using it in any scientific way. I _personally_ just think it is sad that the number one topic of the day has nothing to do with science or DIY Bio but drug legalization.


On Wednesday, October 31, 2012 4:25:35 PM UTC-5, GeneralOya wrote:

Hmmm not on topic of DIYBio huh. Well it's a proven effective cancer medicine, which folks can grow in their home/yard. What more do you want?

You can also make fuel from it.
You can also make textiles from it.
The seeds are effective in reducing cholesterol.
It gives immediate relief to individuals suffering from a number of conditions.

So believe the lies from an era of prohibition fueled racial profiling, or do the research. It's a fabulous plant. I'm glad to see a few folks chiming in regardless of where they stand.

Ryan

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Re: [DIYbio] I am a programmer looking for webapp idea that can be done in 5 weeks!

On Wed, Oct 31, 2012 at 4:25 PM, Sebastian S. Cocioba
<scocioba@gmail.com> wrote:
> All this gtk# sillyness for emguCV in monodevelop is gonna take me some time to get used to.

You don't have to use the GUI, last time I checked. I just used the commands.

- Bryan
http://heybryan.org/
1 512 203 0507

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Re: [DIYbio] Re: THC legislation is next week

Hmmm not on topic of DIYBio huh. Well it's a proven effective cancer medicine, which folks can grow in their home/yard. What more do you want?

You can also make fuel from it.
You can also make textiles from it.
The seeds are effective in reducing cholesterol.
It gives immediate relief to individuals suffering from a number of conditions.

So believe the lies from an era of prohibition fueled racial profiling, or do the research. It's a fabulous plant. I'm glad to see a few folks chiming in regardless of where they stand.

Ryan

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Re: [DIYbio] I am a programmer looking for webapp idea that can be done in 5 weeks!

All this gtk# sillyness for emguCV in monodevelop is gonna take me some time to get used to. Would you mind if I do it in C# visual 2012 and flush out all the concepts and needs of the app and then port it over to mono? The windows monodevelop pack comes with a migration wizard that converts vs to mono. The pInvoke thing is still an issue in mono but there are work arounds. Just a thought.

Sebastian S Cocioba
CEO & Founder
New York Botanics, LLC

Sent via Mobile E-Mail

On Oct 31, 2012, at 1:23 PM, Nathan McCorkle <nmz787@gmail.com> wrote:

> Sebastian, if you want to do some image processing in .NET or Mono
> using openCV (I've done it before in Visual Studio using an online
> tutorial, it's a PITA more than on linux but it works) I can then move
> the openCV code to C/C++ and throw it under a django or maybe flask
> web GUI (i've been meaning to play with flask).
>
>
> On Mon, Oct 29, 2012 at 12:56 PM, Sebastian S. Cocioba
> <scocioba@gmail.com> wrote:
>> I solely do .NET programming so my offer was for any windows users. The
>> reason why its easy is because i already have code for doing object recog
>> with things broken out so a few tweaks later and it fits. Only catch is that
>> it will be a visual studio project and whatnot. I do want to switch over to
>> more cross platform methods like direct openCV use in C++. Sry for any
>> confusion.
>>
>>
>> Sebastian S Cocioba
>> CEO & Founder
>> New York Botanics, LLC
>>
>> Sent via Mobile E-Mail
>>
>> On Oct 29, 2012, at 3:49 PM, Bryan Bishop <kanzure@gmail.com> wrote:
>>
>> On Mon, Oct 29, 2012 at 2:45 PM, Sebastian S. Cocioba <scocioba@gmail.com>
>> wrote:
>>>
>>> Hmm if anyone would be interested in something like that in a windows app
>>> I could whip it up quickly
>>
>>
>> I am confused.. why is it easier for you to make something that's *not*
>> cross-platform? What?
>>
>> - Bryan
>> http://heybryan.org/
>> 1 512 203 0507
>>
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Re: [DIYbio] I am a programmer looking for webapp idea that can be done in 5 weeks!

Okay, ill slap together a mono project and put it on github as open source. Any suggestions on a cool name for the app? Virtual Cytometer is the working title at the moment. CellSight? Cellenerator? :P

Sebastian S Cocioba
CEO & Founder
New York Botanics, LLC

Sent via Mobile E-Mail

On Oct 31, 2012, at 11:40 AM, Cathal Garvey <cathalgarvey@gmail.com> wrote:

> Was about to suggest this. Meredith tells me that Mono even performs
> better than .NET, so if you already know how to program in .NET,
> consider switching to Mono: it's cross-platform, fast, and open source,
> and you can still program for .NET only at your option. :)
>
> But yea.. would love an application I could run from Linux, and I know a
> lot of the other biohackers on this list and elsewhere favour Linux,
> sometimes Mac; cross platform would be very much appreciated, if you can
> manage it. Thanks for offering!
>
> On 29/10/12 20:48, Bryan Bishop wrote:
>> On Mon, Oct 29, 2012 at 2:56 PM, Sebastian S. Cocioba <scocioba@gmail.com>wrote:
>>
>>> I solely do .NET programming so my offer was for any windows users.
>>
>>
>> Have you heard of the mono CLR? These CLRs are compatible with .NET,
>> ostensibly. Anyway, don't mind me...
>>
>> - Bryan
>> http://heybryan.org/
>> 1 512 203 0507
>
> --
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[**Virtual University Of Pakistan**Student Cafe] .::VULMSIT::.eNoxel.com Urdu Ashaar



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Re: [DIYbio] I am a programmer looking for webapp idea that can be done in 5 weeks!

Sebastian, if you want to do some image processing in .NET or Mono
using openCV (I've done it before in Visual Studio using an online
tutorial, it's a PITA more than on linux but it works) I can then move
the openCV code to C/C++ and throw it under a django or maybe flask
web GUI (i've been meaning to play with flask).


On Mon, Oct 29, 2012 at 12:56 PM, Sebastian S. Cocioba
<scocioba@gmail.com> wrote:
> I solely do .NET programming so my offer was for any windows users. The
> reason why its easy is because i already have code for doing object recog
> with things broken out so a few tweaks later and it fits. Only catch is that
> it will be a visual studio project and whatnot. I do want to switch over to
> more cross platform methods like direct openCV use in C++. Sry for any
> confusion.
>
>
> Sebastian S Cocioba
> CEO & Founder
> New York Botanics, LLC
>
> Sent via Mobile E-Mail
>
> On Oct 29, 2012, at 3:49 PM, Bryan Bishop <kanzure@gmail.com> wrote:
>
> On Mon, Oct 29, 2012 at 2:45 PM, Sebastian S. Cocioba <scocioba@gmail.com>
> wrote:
>>
>> Hmm if anyone would be interested in something like that in a windows app
>> I could whip it up quickly
>
>
> I am confused.. why is it easier for you to make something that's *not*
> cross-platform? What?
>
> - Bryan
> http://heybryan.org/
> 1 512 203 0507
>
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Re: [DIYbio] Re: THC legislation is next week

It's strange that the DEA drug schedule wording uses 'drug or other
substance'. If we take the first defintion for each 'drug' and
'substance':

drug - A substance that has a physiological effect when ingested or
otherwise introduced into the body, in particular.
substance - A particular kind of matter with uniform properties: "a
waxy substance".

http://en.wikipedia.org/wiki/List_of_Schedule_I_drugs_(US)

If a drug is a substance, and a substance has uniform properties then
I definitely don't think Marijuana is a substance, and therefore not a
drug.

If we look at the definition of medicine:
1- The science or practice of the diagnosis, treatment, and prevention
of disease (in technical use often taken to exclude surgery).
2- A drug or other preparation used for the treatment or prevention of disease.

then I would consider it a medicine since that 'other preparation' is there.

Marijuana is a plant that could very well be evolving without us
knowing. Sure the growers or their boss selectively drives this
evolution based on the 'strength' of the resulting product, but
they're definitely not at the end of the line with a GCMS making sure
THC not some new cannabinoid is the reason.

Is this good or bad, I don't know. But as a scientist I know that
natural products can change and vary. They all might get you 'high'
but maybe they don't all affect cancer the same, or neuroprotection or
heavy machinery safety.

Let's not debate safety and health issues though, I've never heard
much good medical research about ethanol other than it being an energy
source/sink and concentrated as a disinfectant.

People like to relax, and not everyone is the same, so I think it's
natural that different neuronal pathways help different people be
comforted or 'in the mood'.

Even using the word 'recreation' in this context is not telling the
whole story, as that word is defined as:
recreation- Activity done for enjoyment when one is not working.

I'm sure there are plenty of people who aren't 'sick' or abusers that
consume medicines to prevent disease.

Disease-
1- A disorder of structure or function in a human, animal, or plant,
esp. one that produces specific signs or symptoms or that affects a...
2- A particular quality, habit, or disposition regarded as adversely
affecting a person or group of people.

So if I know I have to go work on my automobile in the garage, and I
know I will scrape my skin, tear some flesh, get nasty greasy... sure
I'm gonna be drinking beer while I'm at it because beer suppresses
some part of my CNS that responds to pain or the discomfort thereof. I
definitely don't enjoy garage work, so it's not recreation. I guess
that's medical?

On Wed, Oct 31, 2012 at 8:55 AM, Cathal Garvey <cathalgarvey@gmail.com> wrote:
> While I personally agree with your position that drug use is foolish,
> I'm in favour of decriminalisation (I don't say "legalisation") for a
> variety of reasons, both economic and ethical.
>
> Firstly, nonviolent or non-threatening crimes should generally never
> result in jail time. Fines perhaps, community service or psychological
> help, but why incarcerate a person who doesn't pose a threat to anyone
> else? Especially as the research shows that rates of re-offense are
> generally unaffected by jail time.
>
> Secondly that criminalising the drugs provides an avenue for vibrant,
> violence-inspiring drug markets. States should medicalise, not
> criminalise, drug addiction. By "decriminalising" drugs without
> explicitly permitting them for official sale, states can offer free,
> high-purity drug therapy programs to remove any profit margins from drug
> dealers and help remove stigma for addicts.
>
> Thirdly that criminalisation prevents research by forcing would-be
> researchers to undergo expensive and timewasting vetting and control
> paperwork, so research showing what is and isn't dangerous is hard to find.
>
> When it comes to Marijuana, there has actually been some research
> showing that people are *safer* drivers after smoking some. Presumably
> in moderation, of course. I'd be more concerned that, according to some
> admittedly contentious results, THC can trigger conditions in some
> users. Most famously, Parkinson's disease. But to a lesser or more
> anecdotal extent, more frightening conditions such as schizophrenia.
>
> It would seem, and would make sense to me as a geneticist, that one's
> genetic makeup can determine the safety of recreational psychotropics;
> THC is widely known to trigger long-term changes in neural makeup, and
> has even been suggested to trigger nerve regrowth. Well and good in a
> medical context, but nerve regeneration or new neural growth is not
> necessarily a good thing. Your brain spends the first fifth of your
> lifespan pruning neurons *out*, why interfere?
>
> So yea; decriminalise, yes. But I feel it's foolish to fully legalise
> while doubts remain about long-term safety. Decriminalising first would
> allow more real research about safety before taking the step of
> legalising for sale OTC.
> Oddly, this means I'm just as much in favour of decriminalising LSD than
> THC, given the widely known lack of toxicity of the former drug,
> although *obviously* one is far more dangerous to others while tripping
> on LSD than on THC; bad trips, attempting to drive, wandering into
> traffic, etc..
>
> On 31/10/12 14:38, Jeswin wrote:
>> How come there's no test to see if someone if impaired by any drug
>> (similar to alcohol test like breathalyzer or count the alphabet
>> backwards)? If you are impaired while driving any vehicle or while in
>> the public, then you should be arrested for possible injury to others
>> or to self. If found carrying, one should be charged with intent to
>> sell if the amount on person is above a certain limit.
>>
>> I oppose drug use and believe drug abuse/addiction is a
>> medical/psychological condition that needs to be treated by a doctor
>> or related professional.
>>
>> On Wed, Oct 31, 2012 at 10:30 AM, General Oya <generaloya@gmail.com> wrote:
>>> Good question. Here in DC, our nation's capital the voter result was held
>>> from the public at first due to congressional interference and has yet to be
>>> implemented. Although it has been said that medical marijuana will be rolled
>>> out within the year through the liquor stores. In Maryland we've been cowed
>>> under due to our reliance on federal monies. However, Dr. Ben Cardin, the
>>> Democratic senatorial candidate has been the front runner in attempting to
>>> implement medical cannabis for years now. We have a system now, that says
>>> you can get a note from a doctor that can be produced at trial to reduce a
>>> charge to $100 fine, but you still have to go through the hassle of being
>>> arrested and detained and the court battle, while there is nowhere for
>>> citizens to legally acquire their medicines without resorting to the black
>>> market. Prohibition has truly failed and is costing the country dearly.
>>>
>>> Scientists, Doctors, Accountants, Police (L.E.A.P.), Educators, and even
>>> some politicians have become more and more vocal in their support for
>>> legalization/decriminalization.
>>>
>>> I would like to think that the states that have legalization on the ballot
>>> would have enough transparency to assure that the populace's votes are
>>> actually counted and the actual results released. However if one watched the
>>> Fort Collins debate that was shown on National Geographic's American Weed,
>>> you would see that apparently without any increase in turnout the number's
>>> shifted at the last moment in repeal of medical legislation in a very
>>> suspect way.
>>>
>>> In Colorado there is a lab called "Full Spectrum" responsible for testing
>>> the states medicinal strains for THC content and contaminants. I'd love to
>>> bring something like that to Maryland as well.
>>>
>>> Take Care,
>>> Ryan
>>>
>>> On Wed, Oct 31, 2012 at 12:05 AM, Nathan McCorkle <nmz787@gmail.com> wrote:
>>>>
>>>> How do you tell if the laws go through or not?
>>>>
>>>> On Oct 30, 2012 9:04 PM, "GeneralOya" <generaloya@gmail.com> wrote:
>>>>>
>>>>> New PubMed Data published last week on Anti-cancer efficacy of
>>>>> Cannibinoids.
>>>>>
>>>>> http://www.ncbi.nlm.nih.gov/pubmed/23103355
>>>>>
>>>>> Get out there and vote to Legalize!!!
>>>>>
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>>
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Re: [DIYbio] Re: Molecular Dynamics

Protein crystallization has long been known to cause unnatural conformations in proteins due to the proteins been forced to adopt a highly dense rigid body interaction with other proteins in absence of bulk solvent. By unnatural I mean the conformations could perhaps be on the tail end of a Boltzmann distribution or have little representation in the solution ensemble. This is probably less often the case and the effects are usually more subtle. i.e. crystal contacts causing an alpha helix in the protein to adopt a slightly different 3D positioning than would actually occur.

NMR spectrscopy calculates structures using distance constraints and MD methods in solution to find highly represented conformers in the solution ensemble.
Proteins are inherently dynamic and access a wide range of conformations at room temperature in solution but they are still biasedly viewed as static structures. (See work by Dorothy Kern or Lewis Kay)

Comparing the RMSD progression of a structure over time during an MD simulation allows one to find the equilibrium or best fit relaxed states. Do these differ when starting from an X-ray or NMR and if so how.

On Wednesday, October 31, 2012 11:39:26 AM UTC-5, Nathan McCorkle wrote:

On Wed, Oct 31, 2012 at 6:44 AM, Josiah Zayner <josiah...@gmail.com> wrote:
> I was doing some basic MD. I wanted to compare simulations of proteins that
> start from X-ray structures or NMR structures. One would predict that
> because of the nature of crystals that some X-ray structures might simulated
> differently due to starting in unnatural conformation. However, people think
> that X-ray crystallography is the gold standard in structural biology,
> predicting structure &c. when some of these structures are probably
> slightly, majorly off. I need to redo it though actually using NMR and X-ray
> structures from the same proteins instead of different proteins.

Are you saying that sometimes the way proteins crystallize isn't how
they are in situ, and you want to compare many different
crystallization methods with the structures produced to find your way
back to what they're really like in situ? Which you can then compare
with simulations?

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Re: [DIYbio] Re: THC legislation is next week

On Wed, Oct 31, 2012 at 7:38 AM, Jeswin <phillyj101@gmail.com> wrote:
> How come there's no test to see if someone if impaired by any drug
> (similar to alcohol test like breathalyzer or count the alphabet
> backwards)?

I'm sure we'll get to this point in 10 years if we want to (or if it
means more arrests). I could see offset raman spectroscopy possibly
doing it, looking into an eye capillary at the blood in your
head/brain. Urine tests can already do this, and there is research
into MEMS mass spectrometers, but an eye picture is quicker than
asking someone to pee... 'hey I just relieved myself, you'll have to
wait a while'

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Re: [DIYbio] Re: Molecular Dynamics

On Wed, Oct 31, 2012 at 6:44 AM, Josiah Zayner <josiah.zayner@gmail.com> wrote:
> I was doing some basic MD. I wanted to compare simulations of proteins that
> start from X-ray structures or NMR structures. One would predict that
> because of the nature of crystals that some X-ray structures might simulated
> differently due to starting in unnatural conformation. However, people think
> that X-ray crystallography is the gold standard in structural biology,
> predicting structure &c. when some of these structures are probably
> slightly, majorly off. I need to redo it though actually using NMR and X-ray
> structures from the same proteins instead of different proteins.

Are you saying that sometimes the way proteins crystallize isn't how
they are in situ, and you want to compare many different
crystallization methods with the structures produced to find your way
back to what they're really like in situ? Which you can then compare
with simulations?

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Direct Client Need....... QA Analyst with Selenium and Load Runner

Hi ,

 

This is Sarat Kumar from Xperttech Inc.

Please send resumes to saratk@xperttech.com

Our client is looking for a QA Analyst. I am forwarding the requirement  please go through it and let me know your availability and interest.

 

Position: QA Analyst (Selenium and Load Runner is Must)

Location: Cambridge, MA

Duration: 6 months plus

 

Load Runner experience, Advanced selenium

 

Local first….face to face interview is required.

 

·         Advanced selenium (RC or Webdriver) user

·         2+ Load Runner experience

·         3-4 years in QA roles

 

Thanks & Regards,

 

Sarat Kumar

Technical Recruiter

 

400 W Cummings Park,

Suite#2850

Woburn, MA-01801

Email: saratk@xperttech.com

Phone: 781-312-7716

Fax: 781-207-0709

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 For Job Updates





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Direct Client Need....... QA Analyst with Selenium and Load Runner

Hi ,

 

This is Sarat Kumar from Xperttech Inc.

Please send resumes to saratk@xperttech.com

Our client is looking for a QA Analyst. I am forwarding the requirement  please go through it and let me know your availability and interest.

Position: QA Analyst (Selenium and Load Runner is Must)

Location: Cambridge, MA

Duration: 6 months plus

 

Load Runner experience, Advanced selenium

 

Local first….face to face interview is required.

 

·         Advanced selenium (RC or Webdriver) user

·         2+ Load Runner experience

·         3-4 years in QA roles

 

Thanks & Regards,

 

Sarat Kumar

Technical Recruiter

 

400 W Cummings Park,

Suite#2850

Woburn, MA-01801

Email: saratk@xperttech.com

Phone: 781-312-7716

Fax: 781-207-0709

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Re: [DIYbio] Re: DNA design questions

That's interesting. LuxG (or fre) was the limiting factor.
Next it could be Decanal (CDE).

Don't know if there is abundant luxAB. I read that this is not really an enzyme, because it reacts and is then junk (1 proper enzyme catalyses 10'000 molecules substrate, as you kno .

 



On Wed, Oct 31, 2012 at 4:36 PM, Cathal Garvey <cathalgarvey@gmail.com> wrote:
I don't *think* that sequences around the length of a normal promoter in
such close proximity will be a big recombination hazard.. but it's
possible. One way to try to address this would be to read up as much as
possible on the promoter, and run BLAST on the promoter (accepting only
close relatives which are still active for comparison), and try to
identify which parts of the promoter are most important for function.
Then, make some changes to the apparently "unimportant" parts of the
promoter.

This is much harder in eukaryotes than bacteria, because promoter
structure is usually less "logical": in E.coli for example, the
consensus promoter for Sigma Factor 70 (the "constitutive")
transcription factor is:
TTGACANNNNNNNNNNNNNNNNNNNTATAATNNNN[CDS]

Whereas in Eukaryotes and Archaea you get common motifs such as the
TATAAA and the B-recognition sequence SSRCGCC* close to the CDS but with
less stringent spacing, and then you can have enhancers and such up to
several kilobases away. Regardless, this means in Eukaryotes, you have a
similar immediate promoter structure consisting of highly important and
unimportant DNA sequences, but unless you understand which is which you
can't as easily edit things to reduce similarity between otherwise
functionally identical promoters. At least, not without potentially
sacrificing promoter strength.

Something to consider: if I recall correctly, the limiting factor in
luminescence is luciferin availability. That is, having the same
promoter strength for LuxAB and LuxCDE should be a lower priority than
having LuxCDE under strong expression, assuming you can provide LuxCDE
with what it/they need to produce lots of Luciferin. Which is:
tetradecanal. Can't recall if LuxCDE is enough to convert Tetradecanol
to Tetradecanal, or would you need another enzyme to catalyse that..I
think not.

* that's IUPAC notation: S means "strong" = G or C, R means puRine = A or G

On 29/10/12 20:00, Andreas Sturm wrote:
> Sounds interesting thanks!!
>
> But if I do take Promoter -LuxAB - Trmnr - Promoter- LuxCDEG- Trn, the
> chance of homologous recombination wil be there.
>
> And I must take the strongest promoter to get any visible light ammount,
> I'd have to take it twice...
>
>
> If I include the viral 2a oligopeptide and *hope* it works, that will
> circumvent the problem with possible weak links?
>
>
>
>
>
> On Mon, Oct 29, 2012 at 6:22 PM, Cathal Garvey <cathalgarvey@gmail.com>wrote:
>
>> The longer the chain, the more chances there are for a weak link to
>> break it.
>>
>> Issues of transcriptional stalling or translational abortion/stalling
>> could end up killing the whole system, and it's hard to debug. At least
>> if you have several smaller fusions, you can try to identify which part
>> of the system is failing and fix just that.
>>
>> I'd suggest several smaller fusion proteins rather than one giant one.
>> Fuse the critical pairs or sets that form "bottlenecks" in the system:
>> LuxAB are cofactors, so fuse them. LuxCDE are steps in a synthetic
>> chain, so fuse them.
>>
>> On 28/10/12 16:50, Mega wrote:
>>> Does anyone by chance know how long an amino acid chain can be without
>>> getting problems with transcription and/or translation?
>>>
>>> Say you want to make a 6kbp long fusion protein  (you clone the genes in
>>> frame and remove the stop codons) .  That would make 2000 amino acids(!),
>>> and those sequences I've seen so far have around 400 amino acids.
>>>
>>>
>>> Will there be problems with the RNA polymerase or during translation?
>>> (Google didn't hit any adequate results, unfortunately)
>>>
>>>
>>>
>>>
>>
>>
>> --
>> www.indiebiotech.com
>> twitter.com/onetruecathal
>> joindiaspora.com/u/cathalgarvey
>> PGP Public Key: http://bit.ly/CathalGKey
>>
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>

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