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
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: diybio@googlegroups.com
Subject: Re: [DIYbio] Re: On the topic of safety and DIY tools
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