Giczey G, Kerényi Z, Dallmann G, Hornok L
Agricultural Biotechnology Center, Gödöllö, Hungary.
FEMS Microbiol Lett. 1998 Aug 15;165(2):247-52. doi: 10.1111/j.1574-6968.1998.tb13153.x.
A 42-kDa endochitinase encoding gene, Tham-ch, was cloned by screening the genomic library of Trichoderma hamatum strain Tam-61 with a PCR-amplified chitinase sequence from the same fungus. Tham-ch with its own regulatory sequences was reintroduced into the host strain. The integration of the transforming construct was stable only in one copy. Homologous integration occurred in nine transformants, while non-homologous integration was detected in one transformant. All but one transformant expressed higher levels of chitinase activity in comparison to the wild-type recipient strain; the maximum level of increase was 5-fold. Duplicating the copy number of the highly conserved approximately 42-kDa endochitinase encoding gene appears to be one potential means by which the biocontrol capability of the Trichoderma species might be improved.
通过用来自同一真菌哈茨木霉Tam-61菌株的PCR扩增几丁质酶序列筛选其基因组文库，克隆了一个编码42 kDa内切几丁质酶的基因Tham-ch。带有自身调控序列的Tham-ch被重新导入宿主菌株。转化构建体仅以单拷贝稳定整合。在9个转化体中发生了同源整合，而在1个转化体中检测到非同源整合。与野生型受体菌株相比，除1个转化体外，所有转化体均表达出更高水平的几丁质酶活性；最大增加水平为5倍。复制高度保守的约42 kDa内切几丁质酶编码基因拷贝数似乎是提高木霉菌种生物防治能力的一种潜在手段。
