Prolipoprotein diacylglyceryl transferase (lgt) is a gene associated with receptor of V.cholerae phage VP3
Objective To study the receptor associated genes of V.cholerae phage VP3,and determine the role of prolipoprotein diacylglyceryl transferase in the process of VP3 infects to V.cholerae. Methods Transponson insertion was used to screen mutants resistant to VP3. Southern blot was used to identify the insertion copy number of transponson,inverse PCR was used to amplify the flanking sequence of insertion site located on the strain chromosome. The amplified production was sequenced and determined the gene of insertion site. Meanwhile,clone the integrated gene which was disrupted,complementation experiment was used to identify the sensibility of mutant to VP3. Results A VP3-resistant mutant was obtained and the insertion was confirmed with a single insertion by Southern blot,using inverse PCR,the structural gene of insert site was amplified as lgt in V.cholerae coding prolipoprotein diacylglyceryl transferase. Complementation experiment confirmed the role of lgt in VP3 infection,it′s VP3 susceptibility was fully restored. Conclusion Prolipoprotein diacylglyceryl transferase plays an important role in the susceptibility of VP3, it suggests an essential enzyme in the receptor synthesis.