广西巴马小型猪CTSK 基因CDS 区克隆与生物信息学分析
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(广西大学动物科学技术学院, 南宁 530001)

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Q95-33

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Cloning and bioinformatics analysis of CTSK CDS region in Guangxi Bama mini-pig
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(College of Animal Science and Technology, Guangxi University, Nanning 530001, China)

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    摘要:

    目的 对广西巴马小型猪组织蛋白酶K(cathepsin K,CTSK)基因CDS 区进行克隆及生物信息学分析。方法 从广西巴马小猪的皮下脂肪中提取总RNA,通过RT-PCR 方法扩增获得CTSK 基因CDS 区序列,并采用相关生物信息学软件对其序列特征、系统进化、编码蛋白结构和理化性质进行分析。结果 广西巴马小型猪CTSK 基因CDS 全长993 bp,编码330 个氨基酸,与NCBI 上公布的普通猪(NM_214302. 1)CTSK 基因序列完全一致。同源性比对结果发现,CTSK 基因在进化过程中具有较高的保守性。广西巴马小型猪CTSK 蛋白具有明显的亲水性区域,在第17-330 氨基酸位置区域存在一条信号肽序列,不存在跨膜结构域,二级结构元件由α 螺旋、β 折叠、无规卷曲和延伸链组成。修饰结构预测发现CTSK 蛋白存在两处N 糖基化位点,亚细胞定位发现其可能主要分布于内质网和细胞核。结论 本研究成功克隆了广西巴马小型猪CTSK 基因CDS 序列并进行分析,为今后进一步深入研究该基因功能和建立相关疾病动物模型提供理论基础。

    Abstract:

    Objective This study aimed to clone the CDS region of cathepsin K (CTSK) from Guangxi Bama mini-pig and analyze its genetic structure with bioinformatics software. Methods Total RNA was extracted from subcutaneous fat tissue of a Guangxi Bama mini-pig. The CTSK gene CDS region sequence was obtained through RT-PCR.Its sequence characteristics, evolution, encoded protein structure and physical and chemical properties were analyzed using relevant bioinformatics softwares. Results The CDS of the CTSK gene of Guangxi Bama mini-pig was 993 bp and encoded 330 amino acids, which was completely consistent with the published CTSK gene sequence of pig (NM_214302. 1) on NCBI. Homology comparison showed that the CTSK gene has been highly conserved during evolution. The CTSK protein of Guangxi Bama mini-pig had an obvious hydrophilic region and a signal peptide sequence in the region of amino acids 17-330, but had no transmembrane structural domain. Structural prediction indicated that there were two n-glycosylation sites in the CTSK protein, and subcellular localization analysis indicated that it was mainly distributed in the endoplasmic reticulum and nucleus. Conclusions In this study, the CDS region sequence of the CTSK gene in Guangxi Bama mini-pig is successfully cloned and analyzed, which provides a theoretical basis for further study on the function of this gene and the establishment of animal models of related diseases.

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黄叶,瞿秋红,奉玲丽,江雨航,朱思燃,张广杰,张其伟,綦文晶,郭亚芬,兰干球.广西巴马小型猪CTSK 基因CDS 区克隆与生物信息学分析[J].中国实验动物学报,2019,27(1):96~103.

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  • 收稿日期:2018-07-03
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  • 在线发布日期: 2019-03-06
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