[Expression in E. coli of protein encoded by human esophageal cancer related gene-1].
OBJECTIVE
To express esophageal cancer related gene-1 (ECRG-1)--in E. coli.


METHODS
The human ECRG-1 was cloned by RT-PCR. Expression plasmid of the ECRG-1/GST pi fusion gene was constructed and introduced into E. Coli. The fusion protein was induced to express by ITPG. The recombinant protein was identified by Western blot. BALB/C mice were immunized with the protein purified by SDS-PAGE for the preparation of polyclonal antibody.


RESULTS
DNA sequencing confirmed that the sequence of ECRG-1 was identical to that of the previously obtained by 5'-RACE technique. Expression of the protein in E. coli was verified by Western blot. The polyclonal antibody obtained from immunized BALB/C mice had a title of 1:3,200.


CONCLUSIONS
The antibody available is useful for the further study of structure and function of the esophageal cancer related protein.
Wang
Y
Y
Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021, China.
Tang
H
H
Lu
S
S
D000818:Animals; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D057057:Serine Proteases; D014162:Transfection
D029968:Escherichia coli Proteins; D007527:Isoenzymes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C446704:TMPRSS11A protein, human; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase; C496557:Gstp1 protein, mouse; D057057:Serine Proteases