Re: [DIYbio] Re: On the topic of safety and DIY tools

-it's alright even though it's mostly my fault.

-Regarding #1 & 2 Quantity yes, what about quality? If what you say is true about them being more bacteria-like, don't you run the risk of lack in solubility due to lack in glycosylation?

-My private matter is not so private anymore. I am running a fundraiser, on experiment.com. Not doing a good job of it I am afraid. Someone told me lack in detail is doing me in. Plus a video would help. You and anyone interested is welcome to poke holes in it. For example gene expression stability always seems to be on your mind. I haven't thought of it yet.

Bore anyone else with plant biotech? If I wouldn't be as volatile, I'd moderate a group devoted to that in DIY bio.

On Friday, July 18, 2014 11:24:39 PM UTC-4, Sebastian wrote:
> Thanks for understanding and truly sorry for the confusion. I can get worked up about silly things and seem hostile. I put my foot in my mouth and for that I am sorry.
>
>
> The main reason Im targeting the chloroplast is simple and two-part:
>
> 1. There are many more chloroplasts that would express my transgene than the one nucleus per cell. In a plant, chloroplasts express the bulk of all the proteins in a plant cell including the large subunit of RuBisCo. Lots of potential to bring your gene to the top of the total soluble protein list in terms of overall quantity.
>
>
> 2. Since the chloroplast (cp) is of prokaryotic ancestry, it can be manipulated as a bacterium which is simpler than a higher organism. Expression in bacteria involve far fewer "moving parts" than the myriad components of a eukaryotic expression scenario. Bacteria, including the plant chloroplast, can express a long chain of genes with just a single promoter which makes circuit design simpler in a few ways.
>
>
> Its a worthwhile endeavor, albeit quite difficult, to transform the chloroplast and the technology to do so is fairly young so there is plenty of room for new discoveries, better ways to skin said cat, etc.
>
> If you need private consultation regarding your project, since im aware of its sensitive nature, feel free to contact me off-list. I'd be happy to give you more info and not bore everyone else with pages and pages of my plant biotech ramblings. :)
>
>
> Sebastian S. Cocioba
> CEO & Founder
> New York Botanics, LLC
> Plant Biotech R&D
>
>
>
> From: Yuriy Fazylov
> Sent: ‎7/‎18/‎2014 10:59 PM
>
> To: diy...@googlegroups.com
>
> Subject: Re: [DIYbio] Re: On the topic of safety and DIY tools
>
>
>
> You know what? You are right. I didn't realize that you mentioned your expression vector in a context of making a kit for gene delivery or a patent on plasmid.
>
> -Why are you so upset and snarky?
>
> My one liners were snarky?
>
>
> Accept my apology and understand that it was difficult to judge a character when he is cool - composed and then flips a switch when he notices you are on the scene asking questions. Understand where the confusion got me. All I did after that was recognize a pattern with you mentioning your expression vector.
>
>
> I had no further interest in your vector after you mentioned how painstaking it was to construct the first time you explained it. Your work is your brainchild. I get that. Don't think that I am bitter about it. I wouldn't even share details of my work if I didn't have to rely on advice from others from time to time.
>
>
> -"publicly salty" what is that? I am afraid I am not familiar with that phrase. Attempts at tracking down the meaning gave me nothing other than cryptography.
>
> -Do you plan on self-experimentation?
> Question built on assumptions but I did work you up to those assumptions so that deserves an answer. Not that I would reserve to self-experimentation but I believe that an inventor should have that freedom. My personal interests are with plants. Most you can do with that for self-experimentation is using its extracts or cook it for dinner. Self-experimentation with a DIY biolistics gun or any such delivery system would be a stupid way to go. There are always other modes of gene delivery and they are getting better. 
>
>
>
> Thanks for the lengthy reply. I had to ask myself, what deserved this kind of reply?
>
> A thought just occurred as I was wrapping this up.
> I am paraphrasing but -bacterial and cp expression is similar. I think I understand why you are targeting cp.
>
>
>
> On Friday, July 18, 2014 6:23:27 AM UTC-4, Sebastian wrote:That is a gross misinterpretation of my post, yuriy. How you could
>
> possibly link my off-list denial of giving you a vector I've made from
>
> scratch for free or otherwise which contains novel, patentable,
>
> elements not found in prior art which I kindly explained to my on-list
>
> post of my rant about someone wanting to do self-experimentation may
>
> hurt themselves because they think its a good idea to use said gene gun
>
> inappropriately is beyond me. How are the two related? Why are you so
>
> upset and snarky? Do you plan on self-experimentation?
>
>
>
> Its quite difficult to convince someone that a rant wasn't intended in
>
> any way towards them but come on now. What did I say that made you
>
> think it had anything to do with you? Im sorry you are upset that I
>
> didn't just hand you months of hard work and heavy financial burden
>
> regardless of compensations offered. I don't think its fair to hold
>
> that against me and twist my words. :( I mean no quarrel, friend and am
>
> happy to help with any and all questions verbosely regarding your
>
> project as I've done previously but I can't just hand out freebies if I
>
> could support myself with them otherwise. Its a moral dilemma and right
>
> now im too broke to throw such plasmid into the public domain yet.
>
>
>
> Please refrain from posting personal issues on the list. Its not fair
>
> to anyone. I would have preferred you just contact me in person and
>
> discuss the issues at hand and not be publicly salty about them. I
>
> don't know how else to convince you that you were, in no way, the
>
> target nor even the after thought of my colorfully worded rant about
>
> bio-punk grinders. Hope we can still maintain a positive, professional
>
> relationship and set this issue aside. Peace, love, science.
>
>
>
> Sebastian S. Cocioba
>
> CEO & Founder
>
> New York Botanics, LLC
>
> Plant Biotech R&D From: Yuriy Fazylov
>
> Sent: ‎7/‎18/‎2014 1:42 AM
>
> To: diy...@googlegroups.com
>
> Subject: [DIYbio] Re: On the topic of safety and DIY tools
>
> Good to know it's not directed at anyone. This coming after I asked
>
> you for an expression vector from you and you just happening to
>
> mention it. No need to hide what you mean.
>
>
>
> Still I am curious about what could do 1500 psi and cost less than
>
> $100. I walked through a hardware store just out of curiosity :)
>
>
>
> I wasn't wrong to ask you for the vector either. Did I noticed a plan
>
> for gene repository here? Nice.
>
>
>
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