Difference between revisions of "Part:BBa K4195091"

Revision as of 12:34, 12 October 2022

myc tag-his-vp0980

myc tag-his-Vp0980 can interact with protein TTPA and TTPB and Myc mouse antibody.

Biology

vp0980

V. Parahaemolyticus transmembrane protein Vp0980 is predicted to harbour four transmembrane regions. Two regions are inside of the membrane and two regions are outside of the membrane. The regions outside of the membrane are likely to specially binds phage tail tubular proteins TTPA and TTPB to mediate phage adsorption(1).

Usage

In order to get the binding ability of purified Vp0980 to purified TTPA or TTPB receptor, a Myc-tag was fused to the N-terminal of his-vp0980. We constructed this part on the expression vector pET-28a(+) by Gibson assembly, then transformed the plasmid into E. coli DH5α & E. coli BL21(DE3). The positive transformants were selected by kanamycin, colony PCR and sequencing.

Characterization

1. Agarose Gel Electrophoresis After transforming the plasmid into E. coli BL21(DE3), colony PCR was used to verify that the plasmid was correct. The expected result was obtained (716 bp).

Fig. 1 The result of colony PCR.

Reference

1.M. Hu, H. Zhang, D. Gu, Y. Ma, X. Zhou, Identification of a novel bacterial receptor that binds tail tubular proteins and mediates phage infection of Vibrio parahaemolyticus. Emerg Microbes Infect. 9, 855-867 (2020). Sequence and Features