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

Abstract

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

CLC Number:

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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Cloning of N-terminal and C-terminal of Pig BLM Gene and Its Expression Analysis in Different Tissues and Diseases

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