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拟穴青蟹DDX5基因的表达及功能初步分析
赵逗裟,姚成杰,张子平,王艺磊
0
(集美大学水产学院、农业农村部东海海水健康养殖重点实验室,福建 厦门 361021;福建农林大学海洋学院,福建 福州 350002)
摘要:
拟穴青蟹(Scylla paramamosain)是我国东南沿海重要的经济养殖蟹类之一。DDX5基因是DEAD-box解旋酶基因家族中的成员,已有研究表明其在生殖细胞的成熟分化及功能维持中起重要的调控作用。我们从拟穴青蟹转录组数据库中筛选获得了DDX5SpDDX5)基因序列,采用实时荧光定量PCR技术分析了其在成熟青蟹各组织、性腺各发育时期及胚胎各发育时期的表达模式。此外,利用RNAi技术分析了SpDDX5在性腺发育中可能发挥的作用。结果如下:SpDDX5的开放阅读框长1 893 bp,编码630个氨基酸,具有DEXDc和HELICc两个结构域。SpDDX5在拟穴青蟹的各组织中广泛表达,且在精巢中的表达水平明显高于其他组织(P<0.05);在精巢发育过程中,SpDDX5的表达水平逐渐降低;在胚胎发育过程中,SpDDX5的表达水平逐渐升高。干扰SpDDX5后,生殖细胞分子标记基因VasaNanos表达上调,性腺发育相关基因foxl-2表达下调,Dmrt家族基因Dsx表达上调,Dmrt-likeidmrt-2表达下调。根据以上结果,我们推测SpDDX5基因可能在拟穴青蟹生殖细胞发育和精巢发育中发挥重要作用。
关键词:  海洋生物学  DDX5基因  DEAD-box解旋酶  RNAi  生殖细胞发育  精巢发育  拟穴青蟹
DOI:10.3969/J.ISSN.2095-4972.20230213001
基金项目:国家自然科学基金(41676161);中央引导地方科技发展专项(2020L3011);福建省海洋经济发展专项(FJHJF-L-2022-18)
Expression and functional analysis of DDX5 gene in Scylla paramamosain
ZHAO Dousha,YAO Chengjie,ZHANG Ziping,WANG Yilei
(Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Fisheries College, Jimei University, Xiamen 361021, China;School of Marine Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China)
Abstract:
Scylla paramamosain is one of the most important cultured commercial crabs on the southeast coast of China. DDX5 gene is a member of the DEAD-box helicase gene family. Previous studies showed that it played an important regulatory role in the maturation, differentiation and function maintenance of germ cells. SpDDX5 gene sequence from the transcriptome database of S. paramamosain was obtained and its expression patterns in various tissues, testicular development and embryo development were examined by qRT-PCR. In addition, the possible role of SpDDX5 in gonadal development was analyzed by the RNAi technique. Results show that the open reading frame of SpDDX5 is 1 893 bp in length, which encodes 630 amino acids and has two domains of DEXDc and HELICc. SpDDX5 was widely expressed in various tissues and the expression level of SpDDX5 in the testis was significantly higher than that of other tissues (P<0.05). During testicular development the expression level of SpDDX5 decreased gradually. During embryonic development the expression level of SpDDX5 gradually increased. After SpDDX5 interference, the expression level of Vasa and Nanos, the molecular marker genes, was up-regulated. The expression level of gonadal development-related foxl-2gene was down-regulated. The expression level of Dsx was up-regulated and Dmrt-like and idmrt-2, the genes from Dmrt family, were down-regulated. According to the results, SpDDX5 gene may play an important role in the development of germ cells and testes of S. paramamosain.
Key words:  marine biology  DDX5 gene  DEAD-box helicase  RNAi  germ cell development  testicular development  Scylla paramamosain

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