Construction of DNA damage response gene pprI function-deficient and function-complementary mutants in Deinococcus radiodurans

Guanjun Gao, Huiming Lu, Lifen Huang, Yuejin Hua

Research output: Contribution to journalArticle

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Abstract

PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radiodurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carrying kanamycin resistance gene with the D. radiodurans groEL promoter was cloned by PCR amplification and reversely inserted into the pprI locus in the genome of the wild-type strain R1. The resulting PprI-deficient strain, designated YR1, was very sensitive to ionizing radiation. Meanwhile, the recombinant DNA fragment was cloned into the shuttle vector pRADZ3, and resulted in plasmid pRADK with kanamycin resistance in D. radiodurans. The fragments containing complete pprI gene and 3′-terminal deletion pprIΔ were cloned into plasmid pRADK. The resulted plasmids designated pRADKpprI and pRADKpprIΔ were then transformed to YR1. Results show that YR1 carrying pRADKpprI was able to fully restore the extreme radioresistance to the same level as the wild-type D. raiodurans R1, whereas YR1 pRADKpprIΔ failed to do so. Construction of DNA repair switch PprI function-deficient and function-complementary mutants in D. radiodurans is not only useful to elucidating the relationship between domains and functions of PprI protein, but also opens the door to the further studies of the biological functions of PprI protein in vivo.

Original languageEnglish (US)
Pages (from-to)311-316
Number of pages6
JournalChinese Science Bulletin
Volume50
Issue number4
DOIs
Publication statusPublished - Dec 1 2005

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Keywords

  • Complementary
  • Deinococcus radiodurans R1
  • Mutants
  • ppri

ASJC Scopus subject areas

  • General

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