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Knocking out Analysis of the CpxP gene using Crispr/Cas9 in Escherichia coli MG1655
  • +2
  • xiaoliang he,
  • Yuwen Ren,
  • Wanli Meng,
  • Xinran Yu,
  • Xiaohui Zhou
xiaoliang he
Hebei University of Science and Technology
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Yuwen Ren
Hebei University of Science and Technology
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Wanli Meng
Hebei University of Science and Technology
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Xinran Yu
Hebei University of Science and Technology
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Xiaohui Zhou
Hebei University of Science and Technology
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Abstract

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.

Peer review status:POSTED

04 May 2020Submitted to Biotechnology and Bioengineering
05 May 2020Assigned to Editor
05 May 2020Submission Checks Completed