文章摘要
文蛤SRBI基因变异与红壳色性状的关联性分析
Mutation in SRBI gene and its association with the red shell color in Meretrix meretrix
投稿时间:2020-10-07  修订日期:2021-03-07
DOI:
中文关键词: 文蛤  红壳色  SRBI  SNP  蛋白表达
英文关键词: Meretrix meretrix  red shellScolor  SRBI  SNP  protein expression
基金项目:国家自然科学基金(31772846);国家重点研发计划“蓝色粮仓”科技创新课题(2018YFD0901404);国家现代贝类产业技术体系项目(CARS-49);浙江省生物工程一流学科(A类)自设课题(ZS2019001)项目资助
作者单位邮编
施鹤鸣 浙江万里学院 315100
董迎辉 浙江万里学院 315000
姚韩韩 浙江万里学院 
包永波 浙江万里学院 
林志华 浙江万里学院 
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中文摘要:
      为了解文蛤类胡萝卜素代谢相关基因清道夫受体B类I(Mm-SRBI)与红壳色形成的关联性,采用直接测序法对基因编码区进行SNP筛查和关联分析,并用免疫荧光、蛋白免疫印迹分析了Mm-SRBI蛋白在红、白壳色文蛤外套膜中的表达差异。结果显示,在Mm-SRBI基因的编码区筛选到15个SNP位点,位于保守序列上的4个突变位点c.647C/T、c.723A/G、c.818C/T、c.1037A/G与红、白壳色存在显著关联,其中c.723A/G位点为非同义突变(异亮氨酸突变为缬氨酸),且4个位点在遗传上存在强连锁性;蛋白免疫印迹与免疫荧光结果表明,Mm-SRBI在红壳中的蛋白表达量显著高于白壳文蛤,主要表达部位为外套膜边缘膜的纤毛处。本研究表明,Mm-SRBI基因的高表达与变异可能导致文蛤对类胡萝卜素的代谢产生差异,从而在生长发育过程中导致红壳色的形成。
英文摘要:
      In order to understand the mutation of scavenger receptor class B type I (SRBI) and its association with the formation of red shell color in Meretrix meretrix, we analyzed the SNPs of Mm-SRBI gene CDS by using direct sequencing. Immunofluorescence (IF) and western blot (WB) were used to analyze the expression characteristics of Mm-SRBI protein in the mantle of two shell color strains. The results showed that a total of 15 single nucleotide polymorphisms (SNPs) were detected, of which 4 SNP loci were significantly associated with shell color. All of these mutation sites were on conserved sequences , in which c.723 A/G site was non-synonymous mutation that lead to amino acid changes Ile723Val, and 4 loci were genetically strongly linked. The results of IF and WB revealed that Mm-SRBI protein was higher expressed in the mantle of red shell clams compared to that of the white shell clams, and was mainly expressed at the outer epithelium of the mantle. Our results suggest that the mutations of Mm-SRBI and the high expression of its protein may lead to differences in the metabolism of carotenoids in M. meretrix, which may lead to the formation of red shell color. It also provided a helpful foundation to explore the molecular mechanisms of carotenoids metabolism underlying shell coloration and could be potentially applied to marker-assisted selection breeding programs for M. meretrix.
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