Difference between revisions of "Part:BBa K596004"

Revision as of 14:20, 1 October 2011

Algae Protein Expression Vector

This Biobrick Plasmid is created from pRbcRL (which is also derived from pBluescriptKS-) by putting biobrick prefix and suffix between HSP70A/RbcS2 promoter and 3'UTR sequences from RbcS2 gene. We used pRbcRL(Hsp196)for the backbone plasmid. We remove luciferase protein coding sequence(which is represented in the figure as crluc)using XhoI/BamHI restriction. Then we ligated with excised plasmid with biobrick prefix and suffix. biobrick prefix and suffix contains four restriction enzymes namely XbaI, EcoRI, SpeI, PstI. This plasmid can be used for protein expression in C.Reinhardtii and for cloning purposes in E.coli. it contains ampicillin for selection in E.coli. F1 origin of replication is for double stranded replication process. HSP70A is a sequence for transcriptional activation that enhances expression of the gene.

References:

1. Schroda M, Blöcker D, Beck CF. The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The Plant journal : for cell and molecular biology. 2000;21(2):121-31. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10743653.

2. Fuhrmann M, Hausherr A, Ferbitz L, et al. Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene. Plant molecular biology. 2004;55(6):869-81. Available at: http://www.ncbi.nlm.nih.gov/pubmed/15604722.

Map:

Sequence and Features