华山松疱锈病的重寄生真菌(深绿木霉)中几丁质酶基因cDNA片段的克隆

Cloning of cDNA Fragment of Chitinase Gene from the Mycoparasite Trichoderma atroviride on Armandii Pine Blister Rust

[目的]分离深绿木霉SS003中的几丁质酶基因,为获得高抗病的转基因植株奠定基础。[方法]采用华山松疱锈病的锈孢子壁作为诱导物,诱导深绿木霉SS003中几丁质酶基因的表达,并通过RT-PCR方法扩增得到几丁质酶基因片段。[结果]锈孢子壁可诱导出高活性(40.17μg/10 min)的几丁质酶,PCR扩增得到了长度为834 bp的特异片段,经序列测定及分析显示该片段为几丁质酶基因片段。[结论]深绿木霉SS003的几丁质酶基因片段的获得,为分离全长基因以及进一步利用该基因生产几丁质酶以及进行基因转化提供了可能。

[Objective] The aim of this study was to isolate chitinase gene from Trichoderma atroviride strain SS003. [Method] With the aeciospore wall of armandii pine blister rust as inducer, chitinase gene was induced to express in Trichoderma atroviride cells. The cDNA fragment of chitinase gene was cloned by RT-PCR approach. [Result] The activity of chitinase induced reached 40.17 μg/10 min; and the specific fragment amplified was 834 bp in length and proved to be the fragment of chitinase gene by sequencing and sequence analysis. [Conclusion] The result showed the feasibility of isolating the full length gene and its transformation, and further producing chitinase.