文献类型：期刊文献

中文题名：三角帆蚌dPGAM基因的克隆与表达分析

英文题名：Molecular Cloning and Expression of dPGAM Gene From Hyriopsis cumingii

作者：应晨怡[1];谢陈南[1];张根芳[2];周淼[3];许海方[1];黄芸洁[1];陈旭旭[1];杨受保[1]

机构：[1]绍兴文理学院生命科学学院,浙江绍兴312000;[2]金华职业技术学院农业与生物工程学院,浙江金华321007;[3]诸暨市水产技术推广站,浙江诸暨311800

年份：2019

卷号：0

期号：10

起止页码：1913

中文期刊名：核农学报

外文期刊名：Journal of Nuclear Agricultural Sciences

收录：CSTPCD、、北大核心2017、CSCD2019_2020、北大核心、CSCD

基金：绍兴文理学院中青年学术骨干培养对象(2014);“十二五”浙江省水产育种科技专项(2012C12907-5);浙江省自然科学基金(LY12C19010);国家自然基金(31302210)

语种：中文

中文关键词：三角帆蚌;磷酸甘油酸变位酶;克隆;表达

外文关键词：Hyriopsis cumingii;phosphoglycerate mutase;clone;expression

中文摘要：为探究贝类磷酸甘油酸变位酶(PGAM)基因的功能,以紫色和黄色家系三角帆蚌为试验材料,采用RACE-PCR法克隆得到三角帆蚌1个辅因子依赖型磷酸甘油酸变位酶基因(dPGAM)的cDNA序列,命名为HcdPGAM,并对其进行生物信息学和表达特性分析。结果表明,HcdPGAM基因包含1个750 bp的开放阅读框,编码1个含250个氨基酸的假定蛋白,并含有1个保守的组氨酸磷酸酶结构域;HcdPGAM与其他动物PGAM的氨基酸序列均具有较高的相似性(58.8%~82.0%)。实时荧光定量PCR分析显示,HcdPGAM在所检测的紫色蚌和黄色蚌的鳃、外套膜、闭壳肌、消化腺、性腺组织和血淋巴细胞6种组织中均有表达,其中在闭壳肌中的表达量相对较高,且紫色系蚌闭壳肌中的表达水平显著高于黄色系,表明该基因与三角帆蚌的能量代谢与生长发育有关,是贝类分子辅助育种的一种潜在的标记;嗜水气单胞菌诱导可极显著上调HcdPGAM基因的表达水平(24 h和48 h),表明HcdPGAM基因在贝类抗细菌感染的免疫反应相关能量代谢过程中也发挥着重要作用。本研究结果为明确贝类PGAM的功能和筛选三角帆蚌不同颜色家系间的特异性分子标记提供了一定的理论依据。

外文摘要：In order to explore the function of Phosphoglycerate mutase(PGAM)gene in mollusks,a cofactor-dependent PGAM(named McdPGAM)was cloned by RACE-PCR method from individuals of purple and yellow genus of Hyriopsis cumingii.Its sequence characteristics and expression pattern were analyzed.The results showed that HcdPGAM gene contained a 753 bp open reading frame,which encoded a protein of 250 amino acids with a highly conserved histidine phosphatase domain(4-250 aa).HcPGAM protein shared high homology with PGAM from other animals(58.8%-82.0%).RT-qPCR analysis showed that HcdPGAM was expressed in all of the six examined tissues from the purple-mussels and yellow-mussels,relative higher transcription level was detected in adductor muscle tissue,and significantly higher expression level was detected in the purple-mussels than that of yellow ones,which indicating that HcdPGAM was related to the energy metabolism,growth and development of H.cumingii,and it was a potential marker for selected breeding of mollusks.Moreover,the induction of Aeromonas hydrophila significantly up-regulated the expression level of HcdPGAM gene(at 24 h and 48 h),demonstrating that HcdPGAM gene was also playing an important role in the immune-related energy metabolism of mollusks against bacterial infection.The results of present study providing basic information for further study on the roles of PGAM,and for screening specific molecular markers between different color pedigrees of pearl mussel H.cumingii.