摘要： 生物体中接受蓝光信号的因子为蓝光受体蛋白。为了揭示蓝光受体蛋白在蛹虫草中的存在形式，将蛹虫草蓝光受体基因Cmwc-1和Cmwc-2的部分序列构建在原核表达载体pET-41a （+）中，在大肠杆菌中进行表达。以所表达的融合蛋白作抗原制备抗体，并由Western Blotting对蛹虫草菌丝体和子实体分别进行分析检测。结果表明，获得了高效价的蛹虫草蓝光受体蛋白CmWC-1和CmWC-2的抗体。经过Western Blotting分析，在蛹虫草菌丝体和子实体中检测到CmWC-1有42 ku （CmWC1-42）和48 ku （CmWC1-48）2种蛋白质存在形式，其中在菌丝体中主要以CmWC1-48为主，CmWC1-42的相对量较少；而在子实体中主要以CmWC1-42为主，没有检测到CmWC1-48。在菌丝体和子实体中检测到CmWC-2有相同的50 ku （CmWC2-50）蛋白质存在形式。结果表明，在CmWC-1和CmWC-2复合物行使生物功能过程中，CmWC-1在菌丝体和子实体中分别产生了差异性降解，而CmWC-2维持稳定。

关键词: 蛹虫草, 蓝光受体蛋白, 抗体制备, Western Blotting分析

Abstract: The blue light receptor protein is the factor that receives blue light signal in living organisms. To reveal the existence form of blue light receptor protein, the partial sequences of the blue light receptor genes Cmwc-1 and Cmwc-2 in Cordyceps militaris were constructed in prokaryotic expression vector pET-41a (+), and the correct expression of Cmwc-1 and Cmwc-2 was carried out in Escherichia coli. Using the expressed fusion protein as antigen to prepare antibodies, we obtained the antibodies of high titer of the C. militaris blue light receptor CmWC-1 and CmWC-2. By Western Blotting analysis, 42 ku (CmWC1-42)and 48 ku (CmWC1-48)protein for CmWC-1 were detected in mycelia and fruiting bodies of C.militaris, which in the mycelium was dominated by CmWC1-48, more than the CmWC1-42 relative quantity, while in the fruiting body was dominated by CmWC1-42, no CmWC1-48 was detected. 50 ku (CmWC2-50)protein for CmWC-2 was detected in both mycelia and fruiting bodies in C.militaris. These results suggested that CmWC-1 degraded differently in mycelia and fruiting bodies, while CmWC-2 remained stable after completing the biological functions of CmWC-1 and CmWC-2 complexes.

Key words: Cordyceps militaris, Blue light receptor protein, Antibody preparation, Western Blotting analysis

中图分类号: 

• Q936