Saturday, 9 April 2016

molecular biology - How to put a gene under the control of a regulable promoter?


Suppose I have a hetrologous (eukaryotic, plant) gene sequence that I want to express in E Coli under the control of a regulable promoter (say the lac promoter induced by IPTG) how exactly does one add the promoter sequence upstream of the gene?



Are there any tools to do this?


e.g. Say this is the gene itself, what sequence modification is needed to put it under the control of the lac promoter? The goal is to induce overexpression of the protein coded for by the gene on addition of IPTG to the medium (if I understand the strategy correctly)


atggctaccg ataatgacag ctctgaaaac cgtcgtatgg gtaattacaa gccgtccatc       60

tggaactacg acttcctgca gtccctggct acccgccaca atatcatgga agagcgccac 120

ttgaaactgg cggagaaact gaaaggccag gtgaagttta tgtttggtgc cccgatggag 180
.................................................................
gagccgttcg tgacgttcaa cctgaacagc gttcgtggtt cccatttctt ttacgagttt 1560


ggtgacggtt tcggtgtgac gaatagctgg accaaggttg acatgaagag cgtcctgatt 1620

gatccgattc cactggatga agaataatga 1650


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