Recently I want to express a multimeric protein from Bacillus subtilis in E.coli,this protein has three subunits and the genes coding them are not continuous with each other but separate by 642 and 3 bp,respectively.When used pTrc99a as a vector expressing the protein in E.coli,whether the interval base will be interrupt the translation?
Hi there,
According to the info you give it can be that the three genes belong to the same operon provided the genes have the same orientation (is there any ORF in the 642bp sequence?). If so the cloning of the fragment containing the 3 genes under a strong promoter should result in the efficient production of the complex.
To Dr.Dominique Liger,there is no other ORFs in the 642bp in the gene that coding the protein
Do you know if each gene is under the control of its proper promoter or if the three genes belong to the same operon (one single promoter in the upstream sequence of the first gene and a RBS sequence associated to each ORF)?
To Dr.Dominique Liger，the three genes are expressed by only one promoter and they are in the same operon in Bacillus subtilis.
Ok then only one transcript from which three independent translations will occur
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