Cloning and Secretion
Expression of Heat-shock Protein 70 Gene of Helicobacter
pylori
LIU Chun-Jie, ZHANG Zhao-Shan*, LI Shu-Qin, HUANG
Cui-Fen
( Beijing Institute of Biotechnology, Beijing 100071, China )
Abstract Heat-shock
protein 70 gene (hsp70)was obtained by PCR method from Helicobacter
pylori chromosomal DNA. Sequencing analysis exhibited that the hsp70
gene isolated from Hp Y2 was highly homologous with the gene encoded
in Helicobacter pylori 26695 and J99, which had been sequenced for
complete genome. The hsp70 gene was recombined in vitro with
fusion secretion expression vector pMAL-p2 and was transformed into E.coli
cells. The E.coli strains, containing hsp70 recombinant
plasmid, expressed a 113 kD fusion protein which accounted for 19.4% of the
total bacterial periplasm protein after the induction with IPTG for 5 h at 30
℃. The expressed fusion protein could react specifically with anti-Helicobacter
pylori rabbit IgG, as proved by Western blot method.
Key words Helicobacter pylori；heat-shock protein 70；cloning；secretion expression
*Corresponding author：Tel,86-10-63834140；Fax,86-10-63833521；e-mail,[email protected]