Cloning of Chaperonin Gene from Acidianus tengchongrensis Using Inverse PCR Method
- VernacularTitle:反向PCR方法克隆腾冲嗜酸两面菌的分子伴侣基因
- Author:
Qing MA
- Publication Type:Journal Article
- Keywords:
Acidianus tengchongensis, Inverse PCR, Chaperonin
- From:
Microbiology
1992;0(04):-
- CountryChina
- Language:Chinese
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Abstract:
DNA fragments of ? subunit and ? subunit were amplified by PCR from the genome of Acidianus tengchongensis with the degenerated primers designed from the consensus amino acid sequence of Sulfolobus chaperonins. The two DNA fragments about 500bp were used as probe for southern hybridization to determine the suitable restriction endonuclease. Restriction endonuclease digested genomic DNA was circularized by self-ligation, and complete sequences of ? subunit and ? subunit were obtained by inverse PCR. The primers oriented in the reversed direction of the usual orientation to amplify the DNA sequence that flank the known region. The complete genes of ? subunit and ? subunit were PCR amplified from genomic DNA using two pairs primers.
