大白猪CDO1基因克隆、生物信息学分析及组织表达定位

doi:10.16431/j.cnki.1671-7236.2023.04.007

Abstract

Abstract: 【Objective】 This study was aimed to clone the cysteine dioxygenase type 1(CDO1) gene and conduct bioinformatics analysis,detect the expression of CDO1 gene in different tissues of Large White pigs,and locate the site of CDO1 in mammary gland,so as to provide the basis for further exploring the regulatory role of CDO1 gene in mammary gland development.【Method】 The full-length sequence of CDO1 gene CDS in Large White pigs was amplified by PCR.The CDO1 gene sequence was ligated with pMD18-T vector to transformed into E.coli DH5α competent cells.After PCR identification and sequencing of the amplified culture solution of positive bacteria,the sequence similarity among different species were compared and the phylogenetic tree was constructed.The bioinformatics analysis of CDO1 protein was carried out using online prediction software.The expression of CDO1 gene in different tissues of Large White pigs was detected by Real-time quantitative PCR,the localization of CDO1 protein in the mammary gland of Large White pigs was detected by immunohistochemistry.【Result】 The full-length sequence of CDO1 gene CDS in Large White pigs was 603 bp,encoding 200 amino acids.There was the highest similarity with the nucleotide sequence of CDO1 gene between Large White pigs and Sus scrofa.The phylogenetic tree results showed that Large White pigs and Sus scrofa were first clustered into a group and were closely related to Macaca mulatta.The molecular mass of CDO1 protein was 23.018 ku,the isoelectric point was 5.98,and the instability coefficient was 33.58 (＜40),which was a stable hydrophilic protein and located in the cytoplasm.CDO1 protein had glycosylation and phosphorylation sites.The secondary structure and tertiary structure model of CDO1 protein were consistent.Tissue expression analysis results showed that the expression of CDO1 gene in liver of Large White pigs was the highest,which was significantly higher than that in other tissues (P＜0.05),it was next expressed in mammary gland,longissimus dorsi muscle and heart,which were significantly higher than that in other tissues except for liver (P＜0.05),the expression of CDO1 gene in duodenum was the lowest.Immunohistochemical results showed that CDO1 was localized in somatic cells,epithelial cells and adipocytes of terminal bud (TEB) of mammary gland in sows.【Conclusion 】 The total length of CDO1 gene CDS in Large White pigs was 603 bp,encoding 200 amino acids,which was highly expressed in heart,liver and mammary gland.CDO1 protein was a stable hydrophilic protein,which was enriched in somatic cells,epithelial cells and adipocytes of TEB of mammary gland in sows.The results provided a reference for further exploring the effect of CDO1 protein on mammary gland development and lactation function of sows.

Key words: Large White pigs; CDO1 gene; cloning; expression; localization

CLC Number:

S813.1

LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.

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Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs

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