摘要
蓝光是很重要的环境因素,可以引起生物体的生理和生化变化;而组氨酸激酶为许多生物体中双组分信号转导系统的重要成分.为了研究蓝光对真菌蛹虫草组氨酸激酶基因(Cordyceps militaris Histidine Kinase,CmHK)表达的影响,从真菌蛹虫草中提取总RNA,设计引物并通过RT-PCR获得蛹虫草CmHK基因的ORF序列,同时进行该基因的序列分析.并应用real-time PCR方法分析检测不同时间蓝光照射下蛹虫草菌丝体中CmHK基因的转录情况.结果获得了CmHK基因的ORF序列.对该基因蛋白质翻译产物CmHK进行的生物信息学分析表明,其中存在着4个HAMP功能域、1个HisKA功能域、1个HATPase功能域和1个REC功能域.将蛹虫草菌丝体进行持续的24 h蓝光照射,采用real-time PCR分析期间的CmHK基因表达,在蓝光照射之初CmHK基因的表达量有明显的下降,随后在蓝光持续的照射下有一个震荡上升,并在16 h时达到高峰,随后快速下降.研究表明蛹虫草的CmHK带有典型的组氨酸激酶特征,蓝光的直接照射可以导致CmHK基因表达水平的改变.
Blue light is a very important environmental factor causing physiological and biochemical changes in organisms. Histidine kinase is an important component in the two-component signal transduction system in many organisms. To determine the expression of histidine kinase gene （CmHK） influenced by blue light in fungus Cordyceps militaris, the CmHK ORF sequence of C. militaris was cloned and analyzed. The total RNA was extracted from fungus C. militaris, and the primers designed to obtain a histidine kinase gene （CmHK） ORF sequences of C. militaris by RT-PCR. The real-time PCR method was used to the CmHK gene expression analysis in the C. militaris mycelia with different blue light irradiation duration. The result confirmed that the CmHK gene ORF sequence was obtained. The bioinformatics analysis for the translation products of CmHK gene showed 4 HAMP functional domains, 1 HisKA functional domain, 1 HATPase domain and 1 REC functional domain in CmHK sequence. Putting C. militaris mycelium into continuous 24h blue light irradiation, and using real-time PCR to analyze the expression of CmHK gene in C. militaris during the blue light irradiation, the results showed that the CmHK gene expression had an obvious decrease in the beginning of blue light irradiation, soon afterwards sustained a concussion rising and reached a peak at 16 h, then decreased rapidly. These results suggested that the CmHK in C. militaris has the typical characteristics of histidine kinase, and that direct exposure to blue light can change the expression level of CmHK gene in C. militaris.
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2014年第1期50-55,共6页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(31060004
31260010)
江西省自然科学基金项目(2010GZN0115)
江西省教育厅科技项目(GJJ11387)资助~~
