Prokaryotic Expression of Blue Light Receptor Genes Cmwc-1 and Cmwc-2 and Detection of Expressed Proteins in Cordyceps militaris

doi:10.7668/hbnxb.2018.04.008

Abstract

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

CLC Number:

Q936

ZHONG Xueqing, YE Dexiao, MAO Yumei, FU Mingjia, YANG Zhihai. Prokaryotic Expression of Blue Light Receptor Genes Cmwc-1 and Cmwc-2 and Detection of Expressed Proteins in Cordyceps militaris[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(4): 53-60. doi: 10.7668/hbnxb.2018.04.008.

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