JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (7): 29-33.doi: 10.6040/j.issn.1671-7554.0.2014.903

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Screening of host protein interacting with early protein GP23 of bacteriophage LSPA1 of Salmonella Paratyphi A

MAO Pujia1,2, FENG Mengdie1, HONG Yu3, XU Zeyang1, ZHAO Jihua3, YANG Hongwen3, SONG Wuzhan3, WANG Jiai2, RAO Xiancai4, HUANG Fen1, JING Shenrong1, ZENG Weikun1,2   

  1. 1. Department of Medical Microbiology, Medical Faculty, Kunming University of Science and Technology, Kunming 650500, Yunnan, China;
    2. Department of Clinical Laboratory, Medical Faculty, Kunming University, Kunming 650214, Yunnan, China;
    3. Department of Nuclear Medicine, Kunming General Hospital of Chengdu Military Command, Kunming 650032, Yunnan, China;
    4. Department of Microbiology, Third Military Medical University, Chongqing 404100, China
  • Received:2014-12-05 Revised:2015-03-23 Online:2015-07-10 Published:2015-07-10

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Abstract: Objective To screen the host protein interacting with early protein GP23 of bacteriophage LSPA1 of Salmonella Paratyphi A (S.Paratyphi A) CMCC50973. Methods The genome of S.Paratyphi A (CMCC50973) was digested by Sau3AⅠand the recovered bands (250 to 1 500 bp) were ligated with pAT plasmid to create the S.Paratyphi A genomic library. gp23, an early gene of phage LSPA1 genome, was ligated with pBT as a bait plasmid (pBT-gp23). The S.Paratyphi A genomic library plasmids and pBT-gp23 were co-transformed into the host bacteria KS. Furthermore, the bacterial two-hybrid was used to screen blue single bacterium. Positive clones were tested by the activity of LacZ. Positive plasmids were extracted, sequenced and analyzed. Results S.Paratyphi A genomic library was successfullyconstructed and its capatity met the requirements of the experiment. Positive rate of the library was about 100%. The gene of positive clone was putative purine permease by sequencing and BLAST. Conclusion Host protein(putative purine permease) may interact with early protein GP23 of bacteriophage LSPA1 of S.Paratyphi A.

Key words: Bacterial two-hybrid, S.Paratyphi A, Phage

CLC Number: 

  • Q939.47
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