摘要
寻找并克隆凡纳滨对虾G蛋白α亚基基因,为对虾的生理调控研究提供理论基础.通过简并PCR和RACE技术获得目的基因的全长cDNA序列.将该基因的编码序列克隆到pCMV5表达载体,通过免疫共沉淀实验和胞内cAMP浓度的测定鉴定该基因表达产物的功能.用半定量RT-PCR和Western blotting确定该基因的表达产物在对虾身体各部位的分布.克隆到凡纳滨对虾的G蛋白短式剪切模式Gαs亚基基因,将其表达产物命名为pvGαs-s.pvGαs-s的序列和功能与其它物种的Gαs具有高度保守性.它在对虾身体各部位存在普遍分布,尤其在脑神经和腮中大量表达,在触角、眼等部位也有适量分布.说明pvGαs-s在对虾生命活动中的重要性,为研究对虾的生理调控奠定了理论基础.
In this study,we aimed at identifying new functional genes encoding G protein alpha subunits from Litopenaeus vannamei and elucidating the critical roles that Gα subunits assume in signal transduction process of shrimp. Degenerate PCR and RACE techniques were employed to achieve the full length of Gα subunit in L. vannamei. Co-immunoprecipitarion analysis and measurement of accumulated cAMP in HEK 293 cells overexpressed with this new Gα subunit were used to identify the basic functions. Semi-quan- titative two-step RT-PCR and Western blotting were employed to detect the distributions of the Gα subunit. The short form of Gα,
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2006年第3期409-414,共6页
Journal of Xiamen University：Natural Science
基金
国家高技术"863"计划项目(2002AA629060)资助
关键词
G蛋白
克隆
凡纳滨对虾
was cloned from L. vannamei, and was thus termed pvGα,-s, pvGα,-s was highly conserved in both primary structures and basic functions. The expression patterns revealed that pvGα,-s was widely expressed in many shrimp tissues,especially enriched in brain and gills,it also obviously existed in antennae and eyes. pvGα,-s may play crucial roles in sensory responses and leaching food for shrimp L. vannamei.
