太行鸡CCNB2基因剪接异构体的克隆鉴定及其在卵泡发育中的表达模式分析

doi:10.16431/j.cnki.1671-7236.2022.10.004

Abstract

Abstract: 【Objective】 The aim of the experiment was to clone different splice isomers of cyclin B2 (CCNB2) gene in chicken,and analyze their bioinformatics and tissue expression.【Method】 Reverse transcription PCR was used to amplify and clone the different splice isomers of CCNB2 gene in Taihang chickens.Partial nucleotide sequences of different splice isomers of CCNB2 gene were compared by bioinformatics software, the amino acid sequence similarity of different splice isomers of CCNB2 in different species was compared, the phylogenetic tree was constructed,and its subcellular localization and secondary structure were predicted.The relative expression of different splice isoforms of CCNB2 gene in ovaries and follicles of Taihang chickens at different developmental stages were analyzed by Real-time quantitative PCR.【Result】 In Taihang chickens,the CCNB2 gene had three splicing isomers:CCNB2-AS1,CCNB2-AS2 and CCNB2-AS3,which encoded 390,383 and 401 amino acids,respectively.The amino acid sequence similarity among the three splices CCNB2-AS1,CCNB2-AS2 and CCNB2-AS3 of Taihang chickens was 97.00%,the three spliceosomes had the highest similarity with Gallus,which were 99.71%,99.71% and 100%, respectively.The three splice isomers were evolutionarily closest to Gallus gallus,followed by Phasianus colchicus.Subcellular localization showed that the three splice isoforms were mainly localized in the cytoplasm.Protein secondary structure analysis showed that the three splice isomers were alpha helix.The relative expression levels of CCNB2-AS1,CCNB2-AS2 and CCNB2-AS3 in the follicles of Taihang chickens at different developmental stages were different,and the expression trends of different splice isomers in the follicles at the same stage were also different. CCNB2-AS1 was highly expressed in pregrade follicles.The expression of CCNB2-AS2 in great white follicles was significantly higher than that in other stages (P＜0.05).The expression of CCNB2-AS3 showed an opposite trend to that of CCNB2-AS1,and was highly expressed in graded follicles.【Conclusion】 In this study,three splice isoforms of CCNB2 gene in Taihang chickens were successfully identified,which were expressed in follicles at all stages,but there were differences in the expression trend in pregrade follicles.

Key words: Taihang chickens; CCNB2 gene; splicing isoforms; bioinformatics analysis; tissue expression

CLC Number:

S831.2

WEI Jiarong, ZHANG Beibei, MA Tenghe, LI Mengxiao, LI Xuenan, WANG Bin, FU Guanhua, WANG Hongna, LIU Chao. Cloning and Identification of Splice Isoforms of CCNB2 Gene and Their Expression Patterns in Follicular Development in Taihang Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3726-3735.

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Cloning and Identification of Splice Isoforms of CCNB2 Gene and Their Expression Patterns in Follicular Development in Taihang Chickens

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