德宏奶水牛PRL基因克隆、分子特征及组织差异表达分析  

Cloning,Molecular Characterization and Tissue Differential Expression Analysis of PRL Gene in Dehong Dairy Buffalo

作　　者：谷丽娇 黄丽鸽 盛丹 范新阳[1] 苗永旺[1] GU Lijiao;HUANG Lige;SHENG Dan;FAN Xinyang;MIAO Yongwang(Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2022年第5期780-789,共10页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(31760659)。

摘　　要：【目的】分离鉴定德宏奶水牛催乳素(prolactin,PRL)基因,并分析其分子特征和组织mRNA表达差异。【方法】采用PCR技术对德宏奶水牛PRL基因完整编码序列(CDS)进行分离鉴定,并利用生物信息学方法和半定量RT-PCR技术分别对水牛PRL进行分子特征以及泌乳期和非泌乳期的组织差异表达分析。【结果】水牛PRL基因CDS长690 bp,编码229个氨基酸。水牛PRL蛋白为亲水性分泌蛋白,无跨膜结构域,含有1个信号肽序列(AA1~30)、5种功能修饰位点和1个prolactin_like结构域(AA32~229),且参与了JAK-STAT和PI3K-AKT-mTOR信号通路。PRL的转录区结构及其编码蛋白的结构和保守结构域在牛科物种中高度相似。水牛PRL基因在所检测的11个组织中均有表达,其中,在乳腺中的表达量呈泌乳期极显著高于非泌乳期(P<0.001)。【结论】水牛PRL与其他牛科物种的PRL功能一致。PRL参与JAK-STAT和PI3K-AKT-mTOR乳蛋白合成信号通路,且水牛PRL在乳腺和肺脏等多种组织中均发挥功能,推测其参与水牛乳蛋白和乳脂合成。[Purpose]To isolate and identify the prolactin(PRL)gene in Dehong dairy buffalo and to analyze its molecular characteristics and the expression of mRNA in different tissues.[Methods]The complete coding sequence(CDS)of PRL gene of Dehong dairy buffalo was isolated and identified by PCR,and the molecular characteristics of buffalo PRL and the differential expression of PRL in lactation and non-lactation tissues were analyzed by bioinformatics and semi-quantitative RT-PCR,respectively.[Results]The CDS of PRL gene of buffalo is 690 bp in length and encoded 229 amino acids.The buffalo PRL protein is a hydrophilic secretory protein with no transmembrane structural domain,containing a signal peptide sequence region(AA1-30),five types of functional modifier sites and a prolactin_like structural domain(AA32-229),and participates in JAK-STAT and PI3K-AKT-mTOR signal pathways.The transcriptional region structure of PRL and the structure and conserved structural domains of its encoded protein were consistent in the species of Bovidae.The buffalo PRL gene is expressed in all 11 tissues detected,and the expression in mammary gland during lactation per-iod is extremely significantly higher than that in non-lactation period(P<0.001).[Conclusion]The function of buffalo PRL is highly consistent with that of other species of Bovidae.The PRL particip-ates in JAK-STAT and PI3K-AKT-mTOR milk protein synthesis signaling pathways,and buffalo PRL plays a role in various tissues including mammary gland and lung,so it is speculated that it parti-cipates in buffalo milk protein and milk fat synthesis.

关 键 词：德宏奶水牛 PRL基因 分子特征 组织差异表达 基因功能 

分 类 号：S823.912[农业科学—畜牧学]