猪SGK家族基因的生物信息学分析及其在猪脂肪组织和细胞中的表达  

Bioinformatics Analysis of Porcine SGK Family Genes and Their Expression in Porcine Adipose Tissues and Adipocytes

作　　者：陈川河 刘嘉莉 张立兰 赵莹 陶聪[1,2] CHEN Chuanhe;LIU Jiali;ZHANG Lilan;ZHAO Ying;TAO Cong(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China;Guangdong Laboratory for Lingnan Modern Agriculture,Guangzhou 510642,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]岭南现代农业科学与技术广东实验室,广州510642

出　　处：《中国畜牧兽医》2022年第9期3310-3320,共11页China Animal Husbandry & Veterinary Medicine

基　　金：岭南现代农业实验室科研项目(NT2021005);中央级公益性科研院所基本科研业务费专项资金(2020-YWF-YB-04、2021-YWF-ZYSQ-03)。

摘　　要：【目的】扩增猪血清和糖皮质激素诱导型激酶(SGK)家族基因并进行生物信息学分析,探索其在猪脂肪组织和细胞中的表达模式。【方法】以藏猪脂肪细胞cDNA为模板PCR扩增SGK家族基因,通过在线工具预测其编码蛋白的理化性质及亚细胞定位;用Mega X软件构建系统进化树;采集30日龄巴马猪心脏、肝脏、脾脏、肾脏、肺脏、背肌、腿肌、颈部脂肪、背部脂肪、腹股沟脂肪、肾周脂肪等组织及7日龄和4月龄猪腹股沟脂肪组织,通过实时荧光定量PCR检测SGK家族基因在猪不同部位组织中的表达;采集30日龄巴马猪腹股沟脂肪组织并分离基质血管成分(SVF)细胞,诱导SVF细胞向白色脂肪细胞分化,通过实时荧光定量PCR检测SGK家族基因及脂肪分化标记基因CCAAT增强子结合蛋白α(C/EBPα)、过氧化物酶体增殖物激活受体(PPARγ)在脂肪细胞中的表达。【结果】SGK 1、SGK 2和SGK 3基因CDS区序列长度分别为1296、1104和1473 bp,分别编码431、367和490个氨基酸;SGK1和SGK2定位于细胞质,SGK3定位于细胞核,三者均为亲水性蛋白,3个蛋白均含有相同基序,保守性高;系统进化树结果表明,猪与牛的亲缘关系最近;SGK 1和SGK 3基因在心脏、肝脏、脾脏、肺脏、肾脏、多种肌肉及脂肪组织广泛表达,SGK 2基因在颈部、背部、腹股沟、肾周脂肪组织中均有较高表达;SGK 1和SGK 2基因在7日龄猪脂肪组织中表达量极显著高于4月龄猪脂肪组织(P<0.01),SGK 3基因在4月龄和7日龄的猪脂肪组织中表达量无显著差异(P>0.05),且SGK 3基因的表达量低于SGK 1和SGK 2基因;与未分化脂肪细胞相比,在分化后的脂肪细胞中SGK 1和SGK 2基因的表达量极显著上调(P<0.01),且SGK 1基因的表达量远高于SGK 2基因,而SGK 3基因的表达量无显著变化(P>0.05)。【结论】SGK家族蛋白具有保守结构域,可能发挥着相似的功能,SGK1和SGK2可能参与调控猪脂肪细胞的分化过程,�【Objective】This study was aimed to amplify porcine serum and glucocorticoid-inducible kinase(SGK)family genes and analysis their bioinformatics,as well as to explore their expression patterns in porcine adipose tissues and adipocytes.【Method】SGK family genes were amplified by PCR using the cDNA of Tibetan pig adipocytes.The physicochemical properties and subcellular localization of the encoded proteins were predicted by online tools.The phylogenetic tree was constructed by Mega X software.The tissues from heart,liver,spleen,kidney,lung,back muscle,leg muscle,cervical fat,backfat,inguinal fat,and perirenal fat of 30-day-old Bama pigs and the inguinal adipose tissues of 7-day-old and 4-month-old Bama pigs were collected.The expressions of SGK family genes in these tissues were detected by Real-time quantitative PCR.The 30-day-old porcine inguinal adipose tissues were collected and stromal vascular fraction(SVF)cells were isolated and differentiated into white adipocytes.The expressions of SGK family genes and adipocyte differentiation marker genes CCAAT/enhance binding protein alpha(C/EBPα)and peroxisome proliferator activated receptor gama(PPARγ)in adipocytes were detected by Real-time quantitative PCR.【Result】The sequence length of the CDS regions of SGK 1,SGK 2 and SGK 3 genes were 1296,1104 and 1473 bp,encoding 431,367 and 490 amino acids,respectively.The SGK1 and SGK2 were localized in cytoplasm,and the SGK3 was localized in nucleus.All three SGK proteins were hydrophilic proteins with the same structural domains and conserved motifs of their encoded proteins.Phylogenetic tree results indicated that pigs were most closely related to cattles.SGK 1 and SGK 3 genes were widely expressed in heart,liver,spleen,lung,kidney,various muscles and adipose tissues.SGK 2 gene was highly expressed in adipose tissues of the cervical fat,backfat,inguinal fat and perirenal fat.The expression of SGK 1 and SGK 2 genes were extremely significantly higher in adipose tissues of 7-day-old than 4-month-old pigs(P<0.01),

关 键 词：猪 SGK家族基因 脂肪沉积 表达分析 

分 类 号：S829.9[农业科学—畜牧学]