家兔MSX2基因的克隆表达及生物信息学分析

doi:10.11843/j.issn.0366-6964.2025.06.017

Abstract

Abstract:

This study aimed to investigate the regulatory role of Msh-homeobox 2 (MSX2) in the proliferation of dermal papilla cells (DPCs) in long-haired rabbits. Skin samples were collected from the backs of 12 healthy 6-month-old long-haired rabbits, and DPCs were isolated and cultured. The coding sequence (CDS) of the MSX2 gene was cloned, and its biological characteristics were preliminarily analyzed using bioinformatics tools. The effects of MSX2 overexpression and knockdown on DPCs proliferation were explored, along with the regulatory roles of related genes. The results showed that the CDS of the MSX2 gene was 807 bp in length, encoding 268 amino acids. Overexpression of MSX2 in DPCs significantly increased the mRNA expression levels of hair follicle development-related genes, including CCND1, EGF, Wnt2, and Hoxc13 (P < 0.01), while significantly or extremely significantly decreased the expression of STAT1, SFRP2, and Wnt5a (P < 0.05 or P < 0.01). Conversely, knockdown of MSX2 exhibited the opposite trend. Additionally, MSX2 significantly promoted DPC proliferation. These findings provide a theoretical foundation for further research on the role of MSX2 in hair follicle development in rabbits.

Key words: long-haired rabbits, MSX2, dermal papilla cell, hair follicle development

CLC Number:

S829.1

HAN Xiaoman, SUN Shaoning, YANG Jie, SHEN Ning, BAO Zhiyuan, CAI Jiawei, ZHAO Bohao, CHEN Yang, WU Xinsheng. Cloning Expression and Bioinformatics Analysis of Rabbit MSX2 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2724-2732.

Figures/Tables 10

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项目Projects	用途Purpose	网址/软件Software
ProtParam^[17]	蛋白质的氨基酸序列	https://web.expasy.org/cgi-bin/protparam/
ProtScale^[17]	亲疏水性	https://web.expasy.org/cgi-bin/protscale/
SOPMA^[18]	蛋白质二级结构	https://npsa-prabi.ibcp.fr/
Swiss-Model^[19]	蛋白质三级结构	https://swissmodel.expasy.org/
NetPhos	蛋白的磷酸化位点	http://www.cbs.dtu.dk/services/NetPhos/
	蛋白的N-糖基化位点	http://www.cbs.dtu.dk/services/NetNGlyc/
STRING数据库^[20]	蛋白交联分析	https://string-db.org/cgi/input.pl
STRING data base	MSX2进化分析	MEGA 5.1

siRNAs	引物序列(5′→3′) Primers sequence
siRNA-MSX2	F：GCUUCUCCGUCCAAAGGCATT; R：UGCCUUUGGACGGAGAAGCTT
siRNA-NC	F：UUCUCCGAACGUGUCACGUTT; R：ACGUGACACGUUCGGAGAATT

基因 Gene	引物序列(5′→3′) Primers sequence	产物长度/bp Products length	退火温度/℃ Tm
MSX2	F: CTGCAAGCCACGTCCCTTTA; R: AGGACAAGTGGTACATGCCG	115	60.6
CCND1	F: GAACGCTACCTTCCCCAGTGCTC; R: CCTCACAGACCTCCAGCATCCAG	103	57.2
EGF	F: TGGAGTAGGCATGGAGCAAG; R: GCCCCGTAAGATGGTCGATG	102	59.4
SFRP2	F: CCAGCCCGACTTCTCCTACAAGC; R: TCCAGCACCTCTTTCATGGTCT	135	57.5
STAT1	F: CCAGAGGCCAGTCTTGTCAC; R: ACGGAAACACTACCTGGGAC	112	60.3
Wnt5a	F: CGCAGGACCTGGTCTACATC; R: CGCTCTGTCTGTACCGTCTT	175	59.9
HOXC13	F: AGAAAGAGTACGCAGCCAGC; R: TCTTCTCTTTGACTCGGCGG	123	60.3
Wnt2	F: AGCCATCCAGGTCGTCATGAACCAG; R: TGCACACACGACCTGCTGTACCC	164	56.3
GAPDH	F: CACCAGGGCTGCTTTTAACTCT; R: CTTCCCGTTCTCAGCCTTGACC	145	53.9

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Cloning Expression and Bioinformatics Analysis of Rabbit MSX2 Gene

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