华北农学报2024，Vol.39Issue(2)：174-183,10.DOI:10.7668/hbnxb.20194563
麦洼牦牛MFSD4A基因克隆及组织表达分析
Cloning and Tissue Expression Analysis of MFSD4A Gene in Maiwa Yak
摘要
Abstract
This study aims to obtain the MFSD4 A gene sequence in Maiwa yak and elucidate its expression patterns in different tissues,analyze the mRNA expression levels of the interacting proteins of MFSD4A,and deter-mine the subcellular localization of the MFSD4A protein,and provide a theoretical basis for further research on regu-lating MFSD4 A in the growth and development of yak testis.RT-PCR was used to obtain the MFSD4 A gene CDS se-quence in Maiwa yak and perform bioinformatics analysis.RT-qPCR was used to detect the expression differences of the MFSD4 A gene in 10 tissues,including heart,liver,spleen,lung,kidney,gluteal muscle,back muscle,fat,testis,and lymph,and the correlation between the interacting proteins and MFSD4A.pEGFP-MFSD4 A fusion plasmid was transfected into Madin-Darby bovine kidney cells(MDBK)to investigate the subcellular localization of the MFSD4A protein.The results indicated that the entire length of the MFSD4 A gene CDS region in the macaque yak was 1530 bp,encoding 509 amino acids with a theoretical isoelectric point(PI)of 8.66.It had 12 transmembrane do-mains and belonged to the alkaline transmembrane protein.RT-qPCR results showed differential expression of MFSD4 A mRNA in various tissues of the macaque yak,with the highest expression in the testis.It was also highly positively correlated with the expression levels of MFSD9,CBY1,INHBA,UNC93A,SVOP,and ID1 in the testis,suggesting that the MFSD4 A gene might be related to the regulation of growth and development of yak testis.Fluo-rescence localization of the pEGFP-MFSD4 A fusion vector transfected into Madin-Darby bovine kidney cells(MD-BK)showed that the protein was mainly located in the nucleus.关键词
牦牛/MFSD4A/基因克隆/互作蛋白分析/亚细胞定位Key words
Yak/MFSD4A/Gene cloning/Interaction protein analysis/Subcellular localization分类
农业科技引用本文复制引用
胡莲,刘益丽,赵迪,王泽宁,江明锋..麦洼牦牛MFSD4A基因克隆及组织表达分析[J].华北农学报,2024,39(2):174-183,10.基金项目
四川省科技计划项目(2021YFYZ0001) （2021YFYZ0001）
西南民族大学研究生创新型科研项目(ZD2022490) （ZD2022490）
四川省科技计划项目(2023YFQ0076) （2023YFQ0076）
四川省自然科学基金(2022NSFSC1665) （2022NSFSC1665）
四川省科技计划项目(2023YFQ0076) （2023YFQ0076）
那曲市科技局重点研发项目(NQKJ-2023-07) （NQKJ-2023-07）
