猪BLM基因N端、C端的克隆及其在不同组织与疾病中的表达分析

doi:10.16431/j.cnki.1671-7236.2018.08.005

摘要/Abstract

摘要：

为探究猪BLM解旋酶N端、C端的结构功能及其在不同组织和疾病中的相对表达量，本研究采用RT-PCR技术扩增从江香猪BLM基因N端和C端，利用生物信息学软件对克隆所获序列进行分析，同时采用实时荧光定量PCR技术分析BLM基因在猪不同组织及病样中的表达差异。结果显示，试验成功克隆了从江香猪BLM基因N端1 025 bp和C端1 300 bp序列，与GenBank中猪BLM基因序列（登录号：NM_001123084.1）同源性为100%；生物信息学软件预测显示，猪BLM基因序列长度为4 316 bp，包含4 280 bp的开放阅读框，编码一个由1 426个氨基酸组成的多肽链；氨基酸序列比对发现，猪BLM基因氨基酸序列与牛的同源性最高（85%）；结构域预测表明，猪BLM基因有DEXDc、HELICc、RQC和HRDC 4个结构域；系统进化树分析表明，猪BLM基因与牛亲缘关系最近；实时荧光定量PCR结果显示，以心脏为对照，BLM基因在从江香猪各组织中均有表达，其中在睾丸中的相对表达量最高，其次为肺脏、肝脏、脾脏、心脏、肾脏、小肠和大肠，在睾丸和肺脏中的表达量均极显著高于其他组织（P<0.01）；BLM基因在患繁殖与呼吸道综合征猪肺脏、患白血病鸡肝脏和患鸭瘟鸭肝脏中的表达水平均极显著或显著高于正常组织（P<0.01；P<0.05），提示BLM基因的变化可能也会导致动物疾病的发生。本试验结果可为后续畜禽BLM解旋酶原核表达载体的构建及其在重大疾病的发生和预防等研究提供理论依据。

关键词: 猪; BLM基因; 序列分析; 组织表达

Abstract:

In order to investigate the structure and function of N-terminal and C-terminal of pig BLM gene and its relative expression in different tissues and diseases,the N-terminal and C-terminal of BLM gene from Congjiang Xiang pig were amplified by RT-PCR method in this study,the sequence of clones was analyzed using bioinformatics softwares.At the same time,the expression of BLM gene in different tissues and diseases of pig was analyzed by Real-time quantitative PCR.The results showed that the N-terminal (1 025 bp) and C-terminal (1 300 bp) sequences of BLM gene from Congjiang Xiang pig were successfully cloned,and the sequence homology with GenBank (accession No.:NM_001123084.1) was 100%;The bioinformatics prediction results showed that the sequence length of pig BLM gene was 4 316 bp,including an open reading frame of 4 280 bp,and the corresponding polypeptide was consisted of 1 426 amino acids.Amino acid sequence comparison result showed that the amino acid sequence of pig BLM gene was the highest homology with cow (85%).The phylogenetic tree analysis showed that the pig BLM helicase had four genetic domains of DEXDc,HELICc,RQC and HRDC;Phylogenetic tree analysis showed that the BLM gene relationship between pig and bovine was the closest;Real-time quantitative PCR result showed that compared with the heart tissue,the expression of BLM gene in Congjiang Xiang pig was detected in all tissues,there was the highest relative expression in testis,followed by the tissues of lung,liver,spleen,heart,kidney,small intestine and large intestine.The expression levels in testis and lung were extremely significantly higher than those in other tissues (P<0.01).The expression of BLM gene in lung of porcine reproductive and respiratory syndrome,liver of chicken leukemia and quail duck was extremely significantly or significantly higher than that in normal tissues (P<0.01;P<0.05).The difference indicated that the change in BLM helicase might also lead to animal diseases.The results provided a theoretical basis for construction of prokaryotic expression vector of livestock and poultry BLM helicase and their occurrence and prevention in major diseases.

Key words: pig; BLM gene; sequence analysis; tissue expression

中图分类号:

王赛楠, 许厚强, 赵佳福. 猪BLM基因N端、C端的克隆及其在不同组织与疾病中的表达分析[J]. 《中国畜牧兽医》, 2018, 45(8): 2076-2085.

WANG Sainan, XU Houqiang, ZHAO Jiafu. Cloning of N-terminal and C-terminal of Pig BLM Gene and Its Expression Analysis in Different Tissues and Diseases[J]. , 2018, 45(8): 2076-2085.

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