通过对粉虱、小菜蛾高效的球孢白僵菌HFW-05 几丁质酶基因的克隆及序列分析,旨在从分子水平上了解HFW-05 菌株的几丁质酶特性.根据已发表的几丁质酶基因序列(EU82835)设计几丁质酶基因全长引物,扩增 HFWBbchit1 全长基因,与大肠杆菌(Escherichia coli)克隆载体 pMD19-T 连接获得含有 HFWBbchit1 全长基因的重组质粒 pMDchit1 并测序.该基因的编码区包括1 07 bp,编码了38个氨基酸,分子量约为36.795 kDa,进行同源性比较分析结果表明:其基因序列与球孢白僵菌菌株 NCIM1216 几丁质酶基因(EU82835)和球孢白僵菌菌株 Bb0062 几丁质酶基因(AY150)的核苷酸同源性最高,同源性达到98%.氨基酸序列与球孢白僵菌菌株 Bb0062 几丁质酶(AAN1259)同源性达到99%.
甄伟, 杜立新, 曹伟平, 王容燕, 宋健, 王金耀, 冯书亮
. 球孢白僵菌HFW-05几丁质酶基因的克隆与序列分析[J]. 华北农学报, 2010
, 25(1)
: 36
-39
.
DOI: 10.7668/hbnxb.2010.01.007
To study characters of chitinase from Beauveria bassiana HFW-05 highly toxic to Bemisia tabaci and Plutella xylostella,the chitinase gene HFWBbchit1 was firstly cloned from Beauveria bassiana HFW-05.According to the published sequences of chit1 genes(EU828354),a pair of primers was designed for full length DNA cloning of HFWBbchit1 gene by PCR.Subsequently,the amplified fragment of HFWBbchit1 gene was inserted into Escherichia coli cloning vector pMD19-T and sequenced.Sequence analysis revealed a DNA sequence with a length of 1 047 bp and encoded an open reading frame consisting of 348 amino acids of 70 kDa peptide.The analysis shows that the gene sequence exhibited 98% identity to that of Beauveria bassiana NCIM1216 chitinase(EU828354)and Beauveria bassiana Bb0062(AY145440).The deduced amino acid sequence exhibited 99% identity to that of Beauveria bassiana Bb0062 chitinase(AAN41259)respectively.
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