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Title: Rare codons caused poor expression of mammalian cell entry (Mce1A) gene cloned from Mycobacterium leprae in Escherichia coli
Authors: Santhosh, Ramachandran Sarojini
Dharmalingam, Kuppamuthu
Keywords: Codon usage;Mce1A protein;Mycobacterium leprae;Overexpression;Rare codons
Issue Date: Jan-2016
Publisher: NISCAIR-CSIR, India
Abstract: Mammalian cell entry gene of mycobacterium helps its entry into epithelial and natural target cells. In order to carry out the functional assay of Mycobacterium leprae Mce1A, the whole gene was cloned in Escherichia coli. An overexpression vector carrying M. leprae mce1A gene, under IPTG inducible T5 promoter, was cloned in E. coli for expression of encoded protein with N-terminal 6xHis-tag. In the absence of IPTG, E. coli cells carrying the mce1A gene grew normally but induction of gene expression led to inhibition of cell growth. Western blot analysis using anti-His HRP conjugate showed full length Mce1A protein expressed in low amount. Deletion of N-terminal region having adjacent arginine rare codons resulted in overexpression of truncated protein as inclusion bodies without inhibiting cell division with size reduction of recombinant protein. However the full length protein poorly expressed without size reduction.
Page(s): 64-68
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.15(1) [January 2016]

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