山羊HABP4基因的克隆、序列分析及功能预测

doi:10.11843/j.issn.0366-6964.2021.012.010

摘要/Abstract

摘要： 旨在克隆简州山羊HABP4基因，鉴定与HABP4互作蛋白的mRNA表达水平并分析其相关性，为进一步探讨HABP4基因在调控山羊细胞凋亡中的作用奠定基础。于四川省简阳大哥大牧业有限公司随机选取10月龄左右，体重为55 kg左右的健康简州大耳羊公羊（16只），清晨空腹屠宰后迅速采集各山羊心、肝、脾、肺、肾、小肠、大肠、瘤胃和胰腺9个组织样本，TRIzol法提取组织总RNA，并反转录cDNA。RT-PCR技术克隆山羊HABP4基因cDNA序列，利用在线软件进行生物信息学分析；利用实时荧光定量PCR （RT-qPCR）技术检测HABP4基因在不同组织中的表达量（n=16）以及分析在胰腺（n=16）组织中与该HABP4蛋白可能存在相互作用的10个蛋白的mRNA表达特性及其相关性。结果，成功扩增山羊HABP4基因1 496 bp，包含5'UTR 61 bp，CDS区1 254 bp，3'UTR 181 bp，编码417个氨基酸残基。HABP4 mRNA在山羊各组织中均有表达，在胰腺中表达水平最高，且与UAB52呈极显著正相关，和RPL32、RPS9呈显著正相关，推测其在细胞定位、新陈代谢、多生物过程及生物过程负调节以及翻译起始、核糖体转录、细胞质代谢等方面发挥着重要作用。本研究成功克隆山羊HABP4基因cDNA全长1 496 bp，其可能与UAB52、RPL32和RPS9呈显著正相关。这些结果也为进一步探讨HABP4在调控细胞凋亡过程中的作用提供了参考数据。

关键词: 山羊, HABP4, 克隆, 表达特性, 功能预测

Abstract: The aim of this study was to clone the HABP4 gene sequence of Jianzhou goats, analyze the proteins predicted to interact with HABP4, and determine their mRNA expression, so as to identify the correlations among different genes. These data will be beneficial for exploring the role of HABP4 gene in regulating cell apoptosis in goat. Sixteen 10-month-old healthy Jianzhou goats, with the average weight of 55 kg, were randomly selected from Sichuan Jianyang DAGEDA Animal Husbandary Co. Ltd.. Nine tissue samples of heart, liver, spleen, lung, kidney, pancreas, large intestine, small intestine and rumen of each goat were quickly collected after slaughter. The total RNA was extracted by TRIzol method and reverse transcribed into cDNA. The HABP4 gene sequence was cloned by RT-PCR and sequenced for further bioinformatics analysis using online softwares. Real-time fluorescent quantitative PCR (RT-qPCR) was performed to determine the expression levels of HABP4 in different tissues (n=16), and followed by the analysis of the mRNA expression levels of 10 proteins interacted with the HABP4 protein in pancreatic tissues(n=16). Also, the correlation analysis was performed in expression among these genes. A total length of 1 496 bp HABP4 gene sequence was cloned successfully, including 61 bp of 5' UTR, 1 254 bp of CDS, and 181 bp of 3' UTR, encoding 417 amino acids. The HABP4 expressed in all detected tissues. The mRNA level of HABP4 was higher in pancreas compared with other tissues. The results of correlation analysis showed that the expression of HABP4 was significantly positively correlated with UBA52, RPL32 and RPS9. The results of functional enrichment analysis showed that HABP4 might play an important role in regulating cellular localization, metabolism, negative regulation of biological processes, translation initiation, ribosome transcript and cytoplasmic metabolism. In this study, a full length of 1 496 bp of HABP4 gene was cloned successfully, and which might be significantly correlated with UBA52, RPL32 and RPS9. This data will be beneficial for further exploring the role of HABP4 gene in regulating cell apoptosis.

Key words: goat, HABP4, clone, expression characteristics, functional prediction

中图分类号:

S858.26

王宪军, 向华, 张焕容, 任玉鹏, 朱江江. 山羊HABP4基因的克隆、序列分析及功能预测[J]. 畜牧兽医学报, 2021, 52(12): 3426-3438.

WANG Xianjun, XIANG Hua, ZHANG Huanrong, REN Yupeng, ZHU Jiangjiang. Cloning, Sequence Analysis and Function Prediction of HABP4 Gene in Goat[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3426-3438.

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