琅琊鸡胚MSX2基因克隆及发育性表达变化

doi:10.16431/j.cnki.1671-7236.2022.05.005

Abstract

Abstract: 【Objective】 The aim of this study was to clone Msh-homeobox 2 (MSX2) gene in chickens,and analyze its bioinformatics and embryonic expression patterns,so as to provide support for further study on the structure and function of MSX2 gene.【Method】 The 80 Langya chicken eggs were incubated,the samples were collected on 1st,2nd,3rd,4th,5th,6th,9th,12th,15th and 18th days of incubation,the whole embryos were collected from 1 to 6 embryonic age,the heart and liver were collected from 9,12,15 and 18 embryonic age,respectively).Total RNA was extracted,MSX2 gene was amplified by RT-PCR and cloned,bioinformatics analysis was carried out.Real-time quantitative PCR was used to analyze its expression level in embryo and tissues of Langya chickens.【Result】 MSX2 gene was successfully cloned from Langya chicken embryo with a sequence length of 818 bp,and an open reading frame of 780 bp,encoding 259 amino acids.The MSX2 amino acid sequence of Langya chickens was 99.23% with that of Coturnix japonica,Haliaeetus leucocephalus and Tyto alba alba,and the lowest with that of Capra hircus,which was 74.54%.The phylogenetic tree analysis results showed that Langya chickens and Coturnix japonica was clustered together.Bioinformatics analysis showed that the molecular weight of MSX2 gene was 28 235.18 u,and the isoelectric point (pI) was 9.71.MSX2 protein had no signal peptide and transmembrane domain,and were unstable hydrophilic proteins,and the majority of MSX2 protein was distributed in nucleus (60.9%).The MSX2 protein had 37 phosphorylation sites,8 O-glycosylation sites and 1 N-glycosylation site.The secondary structure of MSX2 protein was mainly composed of random coil (63.32%) and alpha helix (28.19%).Real-time quantitative PCR analysis showed that the expression of MSX2 gene in whole embryo (1-6 embryonic ages) were firstly increased and then decreased,and the expression were the highest at 4 embryonic age,and was extremely significantly higher than 1,2 and 3 embryonic ages (P<0.01).The expression of MSX2 gene in heart of 12 embryonic age were extremely significantly lower than that of 9 embryonic age (P<0.01).However,it showed a gradually decreasing trend in liver of 9-18 embryonic ages,and the expression were the highest in liver of 9 embryonic age (P<0.05).【Conclusion】 The sequence of MSX2 gene in Langya chickens were successfully cloned.The expression patterns of MSX2 gene were different in different embryonic ages and different tissues of the same embryonic age,which provided a foundation for further exploring the structure and function of MSX2 gene in Langya chickens.

Key words: Langya chickens; MSX2 gene; expression; bioinformatics

CLC Number:

S831.2

PANG Yuting, ZHANG Yujie, TANG Weiqi, WANG Yan, XING Jinyi. Cloning and Developmental Expression of MSX2 Gene in Langya Chicken Embryo[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1641-1650.

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Cloning and Developmental Expression of MSX2 Gene in Langya Chicken Embryo

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