User:Silvapy
From 2007.igem.org
Background
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No idea. ask Connie
My name is Laina.
This year I am on the UCB-LBNL IGEM team.
Project
Introduction My task this summer is to isolate and overexpress protochlorophyalide reductase genes (bch-L,-N and -M) and bacteriochlorophyll biosynthetic genes (bch M, N, E, B,and I)of the H. mobilis photosynthetic gene cluster. This is achieved by using upstream ribosome binding sites, downstream from the T7 promoter site, for expression optimization in a high copy number plasmid pET-29a modified to pET29-b-EBBX.
A genomic library was constructed by partial digestion with EcoRI/BamHI and BgII/XhoI followed by ligation of the DA fragments into EcoRI/XhoI abd BgII/XhoI site of the cosmid vector pET29bEBBX. Oligonucleotide primers were synthesized by Intergrated DNA technologies (IDT),Inc, Coraville, IA. Nucleotide sequence analysis was analysed using Lasergene Megasoftware. Database search was done using Basic Local Alignment Search Tool (BLAST). Sequence Compilation Sequence Alignment
File:Bacter1.jpg |
bacteriochlorophyll Biosynthesis Pathway The presence photosynthetic gene cluster involved in the steps of the bacteriochlorophyll/chlorophyll biosynthesis pathway between Mg-chelation and formation of chlorophyllide (bchI, bchD, bchH, bchJ, bchM,bchE,bchL,bchN and bchB) have been found by inspection of the ORFs (Xion et al). |