尼罗罗非鱼pik3r1基因的克隆和mRNA表达分析Cloning Identification and mRNA Expression Analysis of pik3r1 Gene in Oreochromis niloticus
谢彩霞,汪志文,鲁义善,汤菊芬,简纪常
摘要(Abstract)：
【目的】哺乳动物pik3r1基因参与多种免疫途径,探索pik3r1基因在罗非鱼(Oreochromis)中的作用。【方法】克隆尼罗罗非鱼(Oreochromis niloticus)pik3r1(命名为On-pik3r1)cDNA全长,对该基因进行生物信息学分析,并运用荧光定量PCR方法分析无乳链球菌(Streptococcus agalactiae)刺激后On-pik3r1 mRNA在各组织种的表达模式。【结果与结论】On-pik3r1基因编码区2 190 bp,编码729个氨基酸,5′端非编码区(UTR)为542 bp,3′UTR为2 248 bp,理论分子质量为83.99 ku,等电点为5.73。On-pik3r1与斑马拟丽鱼(Maylandia zebra)相似性最高(98.53%),与其他物种的同源性在70%以上,表明pik3r1在物种进化过程中高度保守。On-pik3r1在健康尼罗罗非鱼各组织中均有表达,在肌肉中表达量最高,其次是鳃、皮肤,在胸腺中表达量最低。经灭活无乳链球菌刺激后,On-pik3r1表达量在肠道、鳃、脾脏、头肾、脑部等5个组织中均极显著下调(P <0.01),在胸腺中表现为4 h时极显著下调(P <0.01),24、48、72 h时为显著下调(P <0.05)。On-pik3r1参与了罗非鱼的对无乳链球菌的免疫应答过程。
关键词(KeyWords)： 尼罗罗非鱼;pik3r1;基因克隆;表达;无乳链球菌
基金项目(Foundation): 广东省现代农业产业共性关键技术研发创新团队项目(2019KJ141)
作者(Author): 谢彩霞,汪志文,鲁义善,汤菊芬,简纪常
参考文献(References)：
- [1]张新艳,樊海平,钟全福,等.罗非鱼无乳链球菌的分离、鉴定及致病性研究[J].水产学报, 2008, 32(5):772-779.
- [2]黄锦炉.罗非鱼无乳链球菌病病原学、病理学及cpse基因的原核表达研究[D].成都：四川农业大学, 2012.
- [3]祝璟琳,杨弘,邹芝英,等.海南养殖罗非鱼(Oreochromis niloticus)致病链球菌的分离、鉴定及其药敏试验[J].海洋与湖沼, 2010, 41(4):590-596.
- [4]卢迈新,黎炯,叶星,等.广东与海南养殖罗非鱼无乳链球菌的分离、鉴定与特性分析[J].微生物学通报,2010, 37(5):766-774.
- [5]郭玉娟,张德锋,樊海平,等.中国南方地区罗非鱼无乳链球菌的分子流行病学研究[J].水产学报, 2012,36(3):399-406.
- [6]田珂,贾爱卿,陈善真,等.罗非鱼无乳链球菌生物学特性与致病力研究[J].中国畜牧兽医, 2013, 40(6):203-207.
- [7]柯剑,赵飞,罗理,等.广东省罗非鱼主养区无乳链球菌的分离、鉴定与致病性[J].广东海洋大学学报, 2010,30(3):22-27.
- [8] GEERING B, CUTILLAS P R, NOCK G, et al. Class IA phosphoinositide 3-kinases are obligate p85–p110heterodimers[J]. Proc Natl Acad Sci USA, 2007, 104(19):7809–7814.
- [9]杨佩芳,丁小星,于骏,等. PIK3R1基因及其编码蛋白在宫颈鳞癌中的表达及临床意义[J].江苏大学学报(医学版), 2018, 28(5):423-426.
- [10]林友成. PIK3R1基因在肾细胞癌中的功能及调控机制研究[D].广州：南方医科大学, 2015.
- [11] CHEN Li, YANG Liu, YAO Ling, et al. Characterization of PIK3CA and PIK3R1 somatic mutations in Chinese breast cancer patients[J]. Nature Communications, 2018,9:1357.
- [12]吴迪,刘亚林,王梦昌. PIK3R1对多发性骨髓瘤细胞侵袭转移的影响及其机制初探[J].陕西医学杂志,2015, 44(2):134-137.
- [13] MUNKLEY J, LIVERMORE K E, MCCLURG U L, et al. The PI3K regulatory subunit gene PIK3R1 is under direct control of androgens and repressed in prostate cancer cells[J]. Oncoscience, 2015, 2(9):755-764.
- [14] CHRISTEL Thauvin-Robinet, MARTINE Auclair,LAURENCE Duplomb, et al. PIK3R1 mutations cause syndromic insulin resistance with lipoatrophy[J]. The American Journal of Human Genetics, 2013, 93(1):141-149.
- [15]张丽燕.半滑舌鳎piwil2和pik3r1基因的克隆与表达分析[D].大连:大连海洋大学, 2014.
- [16]聂颖,李昌崇. PI3K信号通路对T细胞发育、活化、增殖、分化及支气管哮喘的作用[J].中华哮喘杂志(电子版), 2011, 5(5):348-352.
- [17] CHEUNG L W T, MILLS G B. Targeting therapeutic liabilities engendered by PIK3R1 mutations for cancer treatment[J]. Pharmacogenomics, 2016, 17(3):297-307.
- [18] STEMKE-KALE K, GONZALEZ-ANGULO A M,LLUCH A, et al. An integrative genomic and proteomic analysis of PIK3CA, PTEN, and AKT mutations in breast cancer[J]. Cancer Res, 2008, 68:6084-6091.
- [19] SHEKAR S C, WU H, FU Z, et al. Mechanism of constitutive phosphoinositide 3-kinase activation by oncogenic mutants of the p85 regulatory subunit[J]. J Biol Chem, 2005, 280:27850-27855.
- [20] TANIGUCHI C M, WINNAY J, KONDO T, et al. The phosphoinositide 3-kinase regulatory subunit p85alpha can exert tumor suppressor properties through negative regulation of growth factor signaling[J]. Cancer Research, 2015, 70(13):5305-5315.
- [21] SAKAI K, SUZUKI H, ODA H, et al. Phosphoinositide3-kinase in nitric oxide synthesis in macrophage critical dimerization of inducible nitric-oxide synthase[J].Journal of Biological Chemistry, 2006, 281(26):17736-17742.
- [22] BOWATER R O, FORBES FAULKNER J,ANDERSON I G, et al. Natural outbreak of Streptococcus agalactiae(GBS)infection in wild giant Queensland grouper, Epinephelus lanceolatus(Bloch),and other wild fish in northern Queensland,Australi[J].Journal of Fish Diseases, 2012, 35(3):173-186.