中国明对虾fenneropenaeus chinensis卵巢及肝胰腺消减文库的构建及分析-construction and analysis of ovarian and hepatopancreas subtractive library of fenneropenaeus chinensis from penaeus chinensis.docxVIP

上海海 上海海洋大学硕士学位论文 Construction and Analysis of Substracted Library in Ovary and Hepatopancreas of shrimp Fenneropenaeus chinensis Abstract Fenneropenaeus chinensis,is an economically important aquaculture species in China.Its taxonomy belong to the phylum Arthropoda,class Crustacea,order Decapoda, family Penaeidae,genus Fenneropenaeus.In the present study,the developmental stages of Chinese shrimp ovarian were determined into five stages:oogonium proliferation stage,previtellogenesis stage,primary vitellogenic stage,secondary vitellogenesis stage and declining stage by external observation,gonad index and paraffin.We performed suppression subtractive hybridization (SSH) experiment and constructed the forward/reverse subtractive library of the ovarian and hepatopancreas from oogonium proliferation stage and primary vitellogenic stage and from previtellogenesis stage and declining stage.The sequencing results were compared using BLASTx homology on GenBank and their functional annotation was conducted via the Gene Ontology.Then we targeted analysis of genes that are differentially expressed during the development.SSH results were validated by the fluorescence quantitative PCR to study the gene expression abundance during the different stages of the ovarian development in order to elucidate the relationship between the expression features and functions of these genes,as well as the relationship between these genes and ovarian development. SSH results show that differentially expressed genes of subtractive library were in accordance with the ovarian development.In the ovarian library,the expression level of differentially expressed genes during the oogonium proliferation was higher in relation with the cellular processes,especially the genes with an inferred function in cell proliferation and differentiation (mitosis).The genes expressed in the stage of previtellogenesis stage,primary vitellogenic stage and declining stage were inferred with the functions of protein synth