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球孢白僵菌蛋白酶成熟因子基因片段的克隆及其表达分析
张军,谢翎,孟慧敏,汪章勋,黄勃
0
(安徽农业大学微生物防治省重点实验室,合肥 230036;安庆师范学院生命科学学院,安庆246011)
摘要:
通过mRNA差异显示技术,从蝉脱诱导培养的球孢白僵菌中分离获得了长度295 bp一种蛋白酶成熟因子的基因片段。该片段编码多肽序列与米曲霉的蛋白酶成熟因子编码氨基酸序列同源性较高,达94.44%。Realtime-PCR检测结果表明,蝉蜕诱导条件下该基因的表达量为对照的8倍左右。诱导下蛋白酶成熟因子的高效表达,表明其在白僵菌毒力相关重组菌株构建研究中可能具有重要的应用价值。
关键词:  球孢白僵菌  蛋白酶成熟因子  mRNA差异显示  荧光定量PCR
DOI:
基金项目:林业公益性行业科研专项(201204506)和农业部公益性行业专项( 201003079)共同资助。
Cloning and expression analysis of a protease maturation factor gene fragment from Beauveria bassiana
ZHANG Jun,XIE Ling,MENG Hui-ming,WANG Zhang-xun,HUANG Bo
(Anhui Provincial Key Laboratory for Microbial Pest Control, Anhui Agricultural University, Hefei 230036;School of Life Sciences, Anqing Teachers College, Anqing 246011)
Abstract:
The fragment of a protease maturation factor gene from Beauveria bassiana was cloned using DDRT-PCR(differential display reverse transcription polymerase chain reaction). The length of the cloned specific fragment was 295 bp. Analysis of the amino acid sequences indicated that the protease maturation factor from Beauveria bassiana was highly homologous to Aspergillus oryzae (94.44%). The expression of the protease maturation factor gene from Beauveria bassiana under different induced conditions was detected using Realtime PCR. We found that the expression of protease maturation factor gene for cuticle of cicada induced was significantly higher than that in control (nearly 8 times of control). It can be inferred that the protease maturation factor gene in Beauveria bassiana may has important application value in construction of genetic engineering strain with high toxicity.
Key words:  Beauveria bassiana  protease maturation factor  DDRT-PCR  Realtime PCR

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