中华绒螯蟹Akt基因的cDNA克隆、序列分析及表达特征
Molecular cloning, sequence analysis and tissue expression of serine/threonine kinases Akt from Eriocheir sinensis
摘要
Abstract
In the present study,full length cDNA encoding the serine/threonine kinases Akt from Eriocheir sinen-sis(EsAkt)was cloned by using 3′RACE and 5′RACE techniques,and the sequence and structural analysis of the EsAkt was conducted with bioinformatics methods. The results showed that the full length cDNA encoding EsAkt consisted of 2 200 bp nucleic acids in length, including a 5′-UTR of 159 bp, a 3′-UTR of 496 bp and an open read-ing frame (ORF) of 1 545 bp encoding 514 amino acids. Analysis of the protein domain features showed that the deduced polypeptides contained three conservative domains characteristic of Serine/Threonine protein kinases family. Multiple sequence alignment revealed that the amnio acids sequences of EsAkt have the 0.889 identity with Fenneropenaeus chinensis and Litopenaeus vannamei.The phylogenetic analysis showed that the EsAkt was ar-ranged in the same clade with Akts from other arthropods.The tissue distribution of EsAkt mRNA in sexual matur-ity individuals and different muscle groups during molt cycle in juvenile crabs were analyzed by quantitative real-time PCR(qRT-PCR).In sexual maturity crabs,the EsAkt transcript was detected in eyestalk,claw muscle,ovary, heart, hepatopancreas, epidermis, testis, gill and triangular membrane, and the expression level was relatively high in ovary,eyestalk and testis,and was low in hepatopancreas.In juvenile crabs,the EsAkt transcript in different muscle groups was different depending on the molt stages.In walking leg muscles,the EsAkt expression level has no obvious change.In abdominal muscles,the EsAkt expression level was much higher in later pre-molt D3?D4 stage than post-molt A-B stage and inter-molt C stage.In claw muscles,the EsAkt expression level was decreased rapidly in pre-molt D3?D4stage and increased in post-molt A-B stage, and lasted to inter-molt C stage. These res-ults suggested that the expression of EsAkt transcript was related with the molt stage of E.sinensis,and it is pos-sible that the EsAkt is involved in the muscle atrophy,growth and rebuilding during the molt cycle of E.sinensis.关键词
中华绒螯蟹/Akt基因/基因克隆/肌肉生长/蜕壳Key words
Eriocheir sinensis/Akt gene/gene clone/muscle growth/molting分类
生物科学引用本文复制引用
田志环,焦传珍,成永旭,吴旭干..中华绒螯蟹Akt基因的cDNA克隆、序列分析及表达特征[J].水产学报,2018,42(4):485-494,10.基金项目
国家自然科学基金(31572635) （31572635）
科技部港澳台科技合作专项(2014DFT30270) （2014DFT30270）
上海市科学技术委员会科研项目(16DZ2281200) （16DZ2281200）
上海高校水产学高峰学科建设项目(2015-62-0908) （2015-62-0908）
上海市科技兴农推广项目[沪农科推字(2015)第1-7号] （2015）
韶关学院生态学重点扶持学科(230079030101)National Natural Science Foundation of China (31572635) （230079030101）
Hongkong, Macao and Taiwan Sci-ence and Technology Cooperation Projects (2014DFT30270) （2014DFT30270）
Projects from Shanghai Municipal Science and Technology Commission (16DZ2281200) （16DZ2281200）
Shanghai Universities First-class Disciplines Project of Fisheries from Shanghai Municipal Education Committee (2015-62-0908) （2015-62-0908）
Agricultural and Technology Promotion Shanghai Mu-nicipal Agriculture Commission (2015D1-7) （2015D1-7）
Key Support Project of Ecology from Shaoguan Univeristy (230079030101) （230079030101）
