#Virulent protein study by cloning and expression
fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR
fbpB–esxA fusing gene ligated by a linker
Clone into expression vector. An expression vector or expression construct is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell.
Express the encoded fusion protein (Ag85B-ESAT-6 fusion protein about 50 kDa ) in Escherichia coli
#Isopropyl ß-D-1-thiogalactopyranoside (IPTG) is a molecular mimic of allolactose, a lactose metabolite that triggers transcription of the lac operon
Identify protein acquired by Western blot using anti-HIS antibody.
Predict the structure and function of the protein
fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR
fbpB–esxA fusing gene ligated by a linker
Clone into expression vector. An expression vector or expression construct is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell.
(https://www.embl.de/pepcore/pepcore_services/cloning/choice_vector/ecoli/vectorfeatures/)
The most commonly used cloning vectors are E. coli plasmids, small circular DNA molecules that include three functional regions: (1) an origin of replication, (2) a drug-resistance gene, and (3) a region where DNA can be inserted without interfering with plasmid replication or expression of the drug-resistance gene.Express the encoded fusion protein (Ag85B-ESAT-6 fusion protein about 50 kDa ) in Escherichia coli
#Isopropyl ß-D-1-thiogalactopyranoside (IPTG) is a molecular mimic of allolactose, a lactose metabolite that triggers transcription of the lac operon
Identify protein acquired by Western blot using anti-HIS antibody.
(http://www.abcam.com/6x-his-tag-antibody-ab137839.html)
Express by N-terminal HIS-tag, and gel filtration to know protein sizePredict the structure and function of the protein
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