高邮鸭Akt1基因克隆及生物信息学分析  

Cloning and bioinformatics analysis of Akt1 gene from Gaoyou ducks

作　　者：李小芬[1] 朱睿 宋易霖 田维婷 张蕾[1] LI Xiaofen;ZHU Rui;SONG Yilin;TIAN Weiting;ZHANG Lei(Jiangsu Agri-Animal Husbandry Vocational College,Taizhou 225300,China;Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development/Jiangsu Key Laboratory for High-Tech Research and Development of Veterinary Biopharmaceuticals,Taizhou 225300,China)

机构地区：[1]江苏农牧科技职业学院,江苏泰州225300 [2]江苏省兽用生物制药高技术研究重点实验室/江苏现代畜牧与新兽药工程技术中心,江苏泰州225300

出　　处：《畜牧与兽医》2024年第10期1-7,共7页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金青年基金项目(32002157);江苏省高校自然科学研究项目(20KJD230001);江苏农牧科技职业学院院级课题(NSFZP202301);2023年度江苏省教育厅“江苏高校‘青蓝工程’”项目(苏教师函[2023]27号);江苏省高职院校青年教师企业实践项目(2023)。

摘　　要：Akt1是丝氨酸/苏氨酸蛋白激酶家族基因,通过响应生长因子刺激磷酸化一系列下游底物来调节许多过程,包括代谢、增殖、细胞存活、生长和血管生成等。实验室前期对全转录组差异分析获得了与高邮鸭双黄蛋率显著相关的候选关键基因Akt1,并且Akt1显著富集于差异表达信号通路——PI3K-AKT信号通路。为了对高邮鸭双黄蛋候选基因Akt1进行系统的功能研究,本研究通过PCR技术克隆高邮鸭卵巢组织Akt1全长,构建了Akt1-EGFP融合表达载体,结合生物信息学技术对AKT1蛋白的理化性质进行系统分析。结果:AKT1为种间保守型非分泌蛋白,表达于质膜,具有丰富的丝氨酸和苏氨酸磷酸化位点,可与10种蛋白(TSC1、MAPK1、PIK3R1、PIK3CA、TSC2、CDKN1A、IKBKB、PIK3CD、PDPK1和ILK)发生互相作用。研究结果为揭示Akt1基因编码的蛋白在调控高邮鸭双黄蛋形成过程中的功能和分子机制提供理论基础。Akt1 is a serine/threonine protein kinase family gene that regulates many processes,including metabolism,proliferation,cell survival,growth,and angiogenesis,by phosphorylating a series of downstream substrates in response to growth factor stimulation.Our research group obtained a candidate key gene,Akt1,that is significantly correlated with the double yolk egg rate of Gaoyou ducks through full transcriptome differential analysis,and Akt1 is significantly enriched in a differentially expressed signaling pathway,the PI3K-AKT signaling pathway.In order to conduct a systematic functional study on the candidate gene Akt1 for the double yolk egg trait of Gaoyou ducks,the full length of the gene in the ovarian tissue of Gaoyou duck was cloned using PCR technology,and an Akt1-EGFP fusion expression vector was constructed.Then,the physicochemical properties of the AKT1 protein were systematically analyzed using bio-informatics technology.The re⁃sults showed that AKT1 was a conserved non-secretory protein between species,expressed on the plasma membrane,with rich serine and threonine phosphorylation sites,and it interacted with 10 proteins(TSC1,MAPK1,PIK3R1,PIK3CA,TSC2,CDKN1A,IKBKB,PIK3CD,PDPK1,and ILK).These results provided a theoretical basis for further exploring the function and molecular mechanism of the protein encoded by the Akt1 gene in regulating the formation of double yolk eggs in Gaoyou ducks.

关 键 词：高邮鸭 双黄蛋 Akt1基因 克隆 生物信息学分析 

分 类 号：S834[农业科学—畜牧学]