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Based on the analysis of  cpxP  genes among  Escherichia coli  strains,  cpxP  gene-targeting short guide RNA (sgRNA) was designed and inserted into the pGL3-MGP-RNA. The donor sequences (MG-HR) for homologous repair were designed and cloned by PCR. MG-HR and pGL3-MGP-RNA were transformed into  E. coli  MG1655 (pCas9). The  cpxP  gene expression cassette was amplified by PCR and subcloned into pBBR1MCS-2. Then the pBBR- cpxP  was independently transformed into  E. coli  MG1655. The results of motility experiment suggest that  cpxP  gene had a significant effect on the movement ability of  E. coli  strain. The CpxP protein had a significant inhibition of bacterial activity. The lastest 81 CpxP proteins sequences were selected and analyzed by multi-sequence alignment and molecular cluster. The CpxP proteins were roughly divided into three categories. Our results suggest that the CpxP protein was involved in bacterial motility, infection and pathogenicity.

Knocking out Analysis of the CpxP gene using Crispr/Cas9 in Escherichia coli MG1655