丽江猪MAPK10基因编码区分离、序列分子特征及组织差异表达分析  

Isolation,Sequence Molecular Characteristics and Tissue Expression of MAPK10 Gene in Lijiang Pig

作　　者：兰国湘[1] 金思琪 严达伟[1] 鲁绍雄[1] 崔艺佳 李国美 李海莲 董新星[1] LAN Guoxiang;JIN Siqi;YAN Dawei;LU Shaoxiong;CUI Yijia;LI Guomei;LI Hailian;DONG Xinxing(Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Xuanwei Livestock and Poultry Improvement Workstation,Yunnan Province,Xuanwei 655400,China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]云南省宣威市畜禽改良工作站,云南宣威655400

出　　处：《云南农业大学学报（自然科学版）》2022年第1期82-90,共9页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(32060736);云南省农业联合重点项目(2018FG001-003)。

摘　　要：【目的】分离丽江猪丝裂原活化蛋白激酶10(mitogen-activated protein kinase,MAPK10)基因序列,并分析其组织mRNA和蛋白表达情况。【方法】采用PCR技术扩增丽江猪MAPK10基因,并利用生物信息学软件对获得的序列进行分析,实时荧光定量PCR检测丽江猪脑、肝脏、心脏、脾脏、肺脏、肾脏、胰脏、背脂和背最长肌等9种组织中MAPK10基因的相对表达量;以大白猪为对照,实时荧光定量PCR和Western-blotting检测丽江猪与大白猪背脂中mRNA和蛋白表达差异。【结果】MAPK10基因序列总长1281 bp,编码426个氨基酸;MAPK10蛋白属亲水蛋白,无跨膜结构和信号肽,有44个磷酸化位点和2个糖基化位点;二级结构以α-螺旋和无规则卷曲为主;丽江猪MAPK10基因序列与人、牛、山羊、绵羊、鼠、狗、猴和鸡的同源性分别为95.3%、93.5%、93.2%、93.0%、92.6%、91.2%、90.5%和85.6%;MAPK10基因在丽江猪9个组织中差异表达,以脑中最高;丽江猪背脂MAPK10基因的mRNA表达量极显著低于大白猪(P<0.01),蛋白表达量也有低于大白猪的趋势(P>0.05)。【结论】MAPK10基因可作为研究丽江猪脂肪沉积性状的候选基因,结果可为研究猪脂肪沉积提供参考。[Purpose]To isolate mitogen-activated protein kinase(MAPK10)gene from Lijiang pig and to analyse the expression of mRNA and protein in tissue.[Methods]The MAPK10 gene in Lijiang pig was amplified by PCR,and the bioinformatics software was used for sequence analysis.The relative expression level of MAPK10 gene in nine tissues of Lijiang pig including brain,liver,heart,spleen,lung,kidney,pancreas,back fat and longissimus dorsi,which was detected by real-time fluorescence quantitative PCR.Taking the Large White pig as a control,real-time fluorescent quantit-ative PCR and Western-blotting were used to detect the difference in mRNA and protein expression in the back fat of Lijiang pig and Large White pig.[Results]The total length of MAPK10 gene se-quence was 1281 bp,which encodes 426 amino acids.MAPK10 protein was a hydrophilic protein without transmembrane domain and signal peptide.There were 44 phosphorylation sites and two glyc-osylation sites.The secondary structure of Lijiang pig MAPK10 was mainly consisted ofα-helices and random coils.The homology between the sequence of Lijiang pig MAPK10 gene and that of hu-man,cattle,goat,sheep,mouse,dog,monkey,and chicken was 95.3%,93.5%,93.2%,93.0%,92.6%,91.2%,90.5%and 85.6%,respectively.MAPK10 mRNA was widely expressed in the nine tissues of Lijiang pigs with highest expression level in brain.The mRNA expression level of MAPK10 gene in back fat of Lijiang pigs was extremely significantly lower than that of the Large White pig(P<0.01),and the protein expression level was also lower than that of the Large White pigs(P>0.05).[Conclusion]MAPK10 gene can be used as a candidate gene for studying fat deposition traits in Lijiang pigs,and the result can provide reference for the study of fat deposition of pigs.

关 键 词：丽江猪 MAPK10基因 生物信息学分析 组织表达 脂肪沉积 

分 类 号：S828.92[农业科学—畜牧学]