乐至黑山羊肾溶菌酶基因克隆及原核表达

Abstract

Abstract: The kidney lysozyme (LZLyK) gene from Lezhi Black goat was cloned, expressed and its activity was detected in this experiment. With the comparative genomics, the LZLyK gene was cloned successfully obtaining a 444 bp long cDNA which coded 147 amino acids. The phylogenetic analysis showed that the LZLyK gene shared the highest homology up to 96.9% with the sheep's lysozyme NO.2(M32498.1) in amino acids. After being transformed into E.coli BL21 (DE3), the recombinant plasmids pET-LZLyK were expressed in a prokaryotic expression system. The SDS-PAGE analysis revealed that the LZLyK protein had molecular mass of 16 and 30 ku respectively, which were consistent with the prediction. The activity of the protein mentioned above was determined by the way of nephelometry, and consequently it turned out to be 15 U/mL which indicated that the protein of LZLyK possessed a certain antibacterial activity. It was found that the content of lysozyme was more in stomach than other tissues through RF-PCR.

Key words: Lezhi Black goat; lysozyme gene; clone; prokaryotic expression

CLC Number:

HU Yu-fei, JIANG Ming-feng, ZHANG Peng, LUO Fan, ZHU Lian-lian, WANG Yong. Cloning and Prokaryotic Expression of Kidney Lysozyme Gene in Lezhi Black Goat[J]. , 2013, 40(9): 46-51.

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Cloning and Prokaryotic Expression of Kidney Lysozyme Gene in Lezhi Black Goat

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