摘要
为研究MAP2K1(MEK1)基因在三角帆蚌(Hyriopsis cumingii)性别决定中的作用,采用cDNA末端快速克隆技术(Rapid-amplification of cDNA ends,RACE)克隆了MAP2K1基因序列,利用实时荧光定量分析比较MAP2K1基因在三角帆蚌6个组织(性腺、闭壳肌、肝胰腺、鳃、外套膜、斧足)、早期发育阶段(1—8月龄)性腺和1—3龄雌雄性腺中的表达水平,利用原位杂交确定MAP2K1基因在2龄三角帆蚌性腺中的定位。结果显示,MAP2K1基因开放阅读框(ORF)长度为1194 bp,编码397个氨基酸。MAP2K1基因在卵巢中高表达;早期发育阶段在2月龄表达量最高;1—3龄的表达结果显示,MAP2K1基因在卵巢中的表达量均高于同期精巢中的表达量(P<0.05)。原位杂交结果显示,MAP2K1基因在雌性三角帆蚌的卵母细胞及卵子上有明显的杂交信号。RNAi结果显示,干扰MAP2K1基因的上游基因C-MOS后,下游基因MAP2K1在雌性中的表达下降了82.31%,在雄性中的表达下降了73.60%。推测MAP2K1基因可能参与三角帆蚌的卵巢发育过程,在三角帆蚌中属于偏雌性基因,其表达受C-MOS基因的影响。
In order to study the role of the MAP2K1(MEK1)gene in the sex determination of Hyriopsis cumingii,we applied RACE(Rapid-amplification of cDNA ends)method to clone the MAP2K1 gene sequence.We conducted a real-time fluorescence quantitative analysis to compare MAP2K1 gene in six tissues(Gonads,adductor muscle,hepatopancreas,gills,mantle,foot),gonads at early developmental stage(1−8 month old)and the 1−3 years'level of expression in male and female glands of H.cumingii.We determined the location of MAP2K1 gene in the gonads of 2-year-old H.cumingii by in situ hybridization.The results show that the ORF region of MAP2K1 gene was 1194 bp in length and encoded 397 amino acids.MAP2K1 gene was highly expressed in the ovary;the expression level was the highest at 2 months of age at early developmental stage;the expression results from 1−3 years of age show that the expression of MAP2K1 gene in the ovary was higher than that in the spermatozoa for the same period(P<0.05).The in situ hybridization results show that the MAP2K1 gene had a significant hybridization signal in the oocytes and eggs of female H.cumingii.RNAi results show that the expression of the downstream gene MAP2K1 gene decreased by 82.31%in females and 73.60%in males after interfering with the upstream gene C-MOS gene of MAP2K1 gene.In conclusion,MAP2K1 gene may be involved in the ovarian development process and is a female-biased gene in H.cumingii,and C-MOS gene affects its expression.
作者
刘美玲
上官宵兆
王晓强
王雅昱
汪桂玲
李家乐
LIU Meiling;SHANGGUAN Xiaozhao;WANG Xiaoqiang;WANG Yayu;WANG Guiling;LI Jiale(Shanghai Ocean University/Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai 201306,China;National Demonstration Center for Experimental Fisheries Science Education/Shanghai Engineering Research Center of Aquaculture,Shanghai 201306,China)
出处
《南方水产科学》
CAS
CSCD
北大核心
2022年第5期91-99,共9页
South China Fisheries Science
基金
国家重点研发计划“蓝色粮仓科技创新”专项(2018YFD0901406)
国家自然科学基金项目(31772835)
财政部和农业农村部:国家现代农业产业技术体系资助。
关键词
三角帆蚌
MAP2K1基因
分子特征
基因表达
Hyriopsis cumingii
MAP2K1 gene
Molecular characterization
Gene expression
