Difference between revisions of "Part:BBa K4195006"

Revision as of 17:32, 10 October 2022

his-vp0980

Biology

Vp0980

V. parahaemolyticus transmembrane protein Vp0980 is predicted to harbour four transmembrane regions, two regions that are inside of the membrane and two regions that 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 and design

In order to certify the interaction between Vp0980 and TTPA/TTPB, we construct this part for immunofluorescence and dot blot. We used both BBa_I0500 to construct the expression system and obtained the composite BBa_K4195110, which are assembled on the expression vector pSB1C3 by standard assembly(Fig. 1). The constructed plasmids were transformed into E. coli BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing. And we characterize the interaction between Vp0980 and TTPA/TTPB.

Fig. 1 Graphic description of the expression gene circuits for display Vp0980 designed in OMEGA project.

Characterization

1. Identification

When constructing this circuit, colony PCR and gene sequencing were used to verify that the transformants were correct. Target bands (2218 bp) can be observed at the position around 750 bp (Fig. 2).

Fig. 2 DNA gel electrophoresis of the colony PCR products of BBa_K4195006_pSB1C3.

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