Re: [DIYbio] Re: DNA design questions

Wikipedia (yeah, not that scientific source) After finding the ribosome binding site in eukaryotes, the ribosome recognizes the Kozak consensus sequence and begins translation at the +1 AUG codon. ( http://en.wikipedia.org/wiki/Ribosomal_binding_site )

Btw,
http://www.invitrogen.com/site/us/en/home/References/Ambion-Tech-Support/translation-systems/general-articles/ribosomal-binding-site-sequence-requirements.html

 Our data demonstrate that in contrast to the E. coli ribosome, which preferentially recognizes the Shine-Dalgarno sequence, eukaryotic ribosomes (such as those found in retic lysate) can efficiently use either the Shine-Dalgarno or the Kozak ribosomal binding sites.

On Sat, Aug 17, 2013 at 8:04 PM, Koeng <koen...@gmail.com> wrote:

I don't think Eukaryotes use RBSs...

http://en.wikipedia.org/wiki/Five_prime_cap

On Saturday, August 17, 2013 6:01:31 AM UTC-7, Mega [Andreas Sturm] wrote:

Hi all,

I am designing primers to get a kanamycin resistance gene.

My question is, the kanamycin gene (nptII)  is out of pGreenII for plants. I want it to work in E.Coli. Will the this eukaryotic RBS also work in E. Coli?

TGACGTTCCATAAATTCCCCTCGGTATCCAATTAGAGTCTCATATTCACTCTCAACTCGATCGAGGCATGATTGAACAA

As I need a BamHI site anyway, I could also mutagenize it into this...

TGACGTTCCATAAATTCCCCTCGGTATCCAATTAGAGTCTCATATTCACTCTCAACagGATCcAGGCATGATTGAACAA

The agga-motive rather looks like prokaryotic beings will accept it?


Or just leave it as it was?

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