白羽番鸭LIF基因克隆及其组织表达差异分析

doi:10.16431/j.cnki.1671-7236.2023.04.002

Abstract

Abstract: 【Objective】 This study was aimed to explore the biological function of LIF gene in White muscovy duck,and clarify its expression pattern in hypothalamus,pituitary,ovary,oviduct enlargement and uterus.【Method】 In this study,the ovarian tissue of White muscovy duck was used as the research object,and the LIF gene sequence of White muscovy duck was cloned by rapid amplification of cDNA ends technology (RACE),and the similarity comparison and phylogenetic tree were constructed.The physicochemical properties and structure of LIF protein were analyzed by bioinformatics methods.The relative expression of LIF gene in the hypothalamus,pituitary,ovary,oviduct enlargement and uterus of White muscovy duck was detected by Real-time quantitative PCR.【Result】 The cloned LIF gene sequence of White muscovy duck was 1 820 bp,and the CDS region was 636 bp,encoding 211 amino acids.The results of nucleotide sequence similarity comparison showed that the sequence similarity of LIF gene between White muscovy duck and Aythya fuligula,Anas platyrhynchos,Oxyura jamaicensis,Cygnus atratus,Serinus canaria, Coturnix japonica,Gallus gallus,Nothoprocta perdicaria were 99.36%,99.21%,99.21%,98.11%,92.03%,90.68%,90.58%,87.60%,respectively.The analysis of phylogenetic tree showed that the genetic distance between White muscovy duck and Aythya fuligula was the closest,and the genetic distance between White muscovy duck and Gallus gallus was the farthest.The LIF protein in White muscovy duck was an alkaline hydrophilic stable protein with 16 phosphorylation sites,7 N-glycosylation sites and 1 signal peptide,which was mainly located in the extracellular and lysosome.The secondary structure of LIF protein was mainly composed of alpha helix and random coli,and the prediction result of protein tertiary structure was consistent with that of secondary structure.LIF protein in White muscovy duck might interact with IL6ST,CNTFR,IL6,OSMR,LIFR,IL6R,STAT3,CNTF,IFNGR1 and IFNGR2 proteins.The tissue expression showed that LIF gene was expressed in five tissues of White muscovy duck. LIF gene of White muscovy duck in the oviduct enlargement and ovarian tissue at nest stage was significantly higher than that at laying stage (P＜0.05).The expression of LIF gene in the oviduct enlargement was significantly higher than other tissues (P＜0.05) in two periods.【Conclusion】 The LIF gene in White muscovy duck was successfully cloned in the experiment,and its expression in the oviduct enlargement was significantly higher than that in other tissues,and the nesting stage was higher than that at laying stage.The results provided a theoretical basis for exploring the regulatory mechanism of ovarian development in White muscovy duck.

Key words: White muscovy duck; LIF gene; bioinformatics; tissue expression

CLC Number:

S834

TAO Qinghua, HUANG Caiyun, CHEN Chao, LIU Wei, SUN Ningning, LI Ang. Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1296-1306.

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Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck

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