绵羊角蛋白关联蛋白2基因的克隆表达与生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.06.004

Abstract

Abstract:

The keratin-associated protein 2 (KAP2) gene was cloned from Xinji Fine-wool sheep and Small-tail Han sheep,the KAP2 proteins was analyzed by bioinformatics,and the relationship between KAP2 gene and molecules trait of wool hair was preliminary analyzed in this study.The total RNA was extracted from wool follicles of Xinji Fine-wool sheep and Small-tail Han sheep.The KAP2 gene was cloned by RT-PCR,and the PCR product was cloned into pMD19-T vector,then was transferred to Escherichia coli DH5α competent cells and the requisite clones were filtered,and the recombinant plasmid was extracted.Identification of PCR and double enzyme digestion and sequencing were operated,then the prediction of protein structure was analyzed by bioinformatics.The results showed that the length of KAP2 gene was 463 bp,containing an ORF that was 414 bp and encoded 137 amino acid residues.The comparison of sequencing results showed that compared with the KAP2 gene in Small-tail Han sheep,there was a base mutation from G to A in KAP2 gene of Xinji Fine-wool sheep which was a missense mutation causing the encoded amino acid changed from arginine (Arg) to glutamine (Gln).The molecular weight of KAP2 protein in Xinji Fine-wool sheep and Small-tailed Han sheep were 14.81 and 14.84 ku,and the isoelectric points of KAP2 protein were 9.67 and 9.82,respectively.The KAP2 protein of the two breeds was no obvious differences in physical and chemical properties,which was transmembrane, alkaline and hydrophobic protein.It was predicted that the secondary structure of KAP2 protein was composed of α-helix,extend chain and random coil.The tertiary structure of KAP2 protein in Xinji Fine-wool sheep and Small-tail Han sheep had a different spatial structure that came from α-helix.Therefore,it could be speculated that KAP2 gene might be a differential gene affecting wool quality.The study of KAP2 gene provided a theoretical basis for exploring the molecular inheritance of phenotypic differences between two wool species.

Key words: keratin-associated protein 2(KAP2); Xinji Fine-wool sheep; Small-tail Han sheep; bioinformatics analysis

CLC Number:

Q785

LIU Xiaojun, ZHANG Lichun, HAN Linkai, CHU Xitong, CUI Chengdu, LU Cheng, JIN Haiguo, SUN Fuliang. Cloning, Expression and Bioinformatics Analysis of KAP2 Gene in Sheep[J]. , 2018, 45(6): 1454-1462.

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Cloning, Expression and Bioinformatics Analysis of KAP2 Gene in Sheep

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