关中奶山羊CSN1S1基因克隆、生物信息学及组织表达谱分析

doi:10.16431/j.cnki.1671-7236.2022.03.007

Abstract

Abstract: 【Objective】 By constructing the tissue expression profile and bioinformatics function of alpha-s1 casein(CSN1S1)gene in Guanzhong dairy goats, the function of CSN1S1 gene in milk composition synthesis of Guanzhong dairy goats was preliminarily investigated.【Method】 Guanzhong dairy goats were taken as the research object, two mutant forms of CSN1S1 were cloned which were named CSNIS1-1 and CSN1S1-2. The protein structure, physicochemical property and phosphorylation sites of CSN1S1 gene and its mutant form were analyzed using a variety of biological information softwares and online tools such as ProtParam, NetPhos, SingalP 4.1 Server, NPS and Phyre2. The relative expression of CSNIS1-1 and CSN1S1-2 in liver, spleen, mammary gland, kidney, uterus and fallopian tube of Guanzhong dairy goats were detected by Real-time quantitative PCR.【Result】 The results showed that two mutant forms CSN1S1-1 and CSN1S1-2 were found in mammary epithelial cells of Guanzhong dairy goats. Sequencing results showed that compared with CSN1S1 sequence, 3 bases mutations were found in CSN1S1-1 and CSN1S1-2, in addition, there were total 6 bases deletions in two positions of CSN1S1-2. The similarity was 99.07% between CSN1S1-1 and CSN1S1, and 97.20% between CSN1S1-2 and CSN1S1.The analysis of protein physicochemical properties showed that the mutation of bases in CSN1S1-1 resulted in the change of leucine at position 31 to proline and glutamine at position 92 to glutamic acid. In addition to the above changes, CSN1S1-2 changed from serine to cysteine at position 83, and changed from glutamic acid to glutamine at position 84. In addition, there were deletions of two serines at positions 81 and 82. Real-time quantitative PCR results showed that the relative expression trends of CSN1S1-1 and CSN1S1-2 in Guanzhong dairy goat tissues were basically the same, and the relative expression levels were higher in mammary gland, uterus and fallopian tube, and basically not expressed in liver, spleen and kidney. The expression of CSN1S1-2 in uterus was significantly higher than that of CSN1S1-1 (P<0.05).【Conclusion】 Two mutant forms CSN1S1-1 and CSN1S1-2 were found in the mammary epithelial cells of Guanzhong dairy goats, and the protein similarity between the two mutant forms and CSN1S1 reached more than 97%. CSN1S1-1 was obviously different from CSN1S1 in protein structure, and the deletion of 6 bases in CSN1S1-2 might be the direct cause of amino acid change, deletion and displacement of phosphorylation site. The expression of CSN1S1-2 in uterus was significantly higher than that of CSN1S1-1, and its potential function in uterus remains to be further explored.

Key words: Guanzhong dairy goats; CSN1S1 gene; cloning; tissue expression profile; bioinformatics analysis

CLC Number:

HOU Jinxing, WANG Zhanhang, ZHU Junru, JIANG Yue, LIU Shujuan, AN Xiaopeng. Cloning, Bioinformatics and Tissue Expression Profile Analysis of CSN1S1 Gene in Guanzhong Dairy Goats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 857-865.

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Cloning, Bioinformatics and Tissue Expression Profile Analysis of CSN1S1 Gene in Guanzhong Dairy Goats

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