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仔猪多胺代谢相关基因的克隆及断奶对其mRNA表达的影响
郭豫杰1,张志强1,王月影1,李宏基1,查光明1,李奎 2,杨国宇1
1. 河南农业大学农业部动物生化与营养重点实验室
2. 中国农业科学院北京畜牧兽医研究所
Molecular Cloning and mRNA Expression Analysis of Polyamine Metabolism-related Genes in Different Tissue of Weaning Piglets (Sus scrofa)
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摘要 多胺是一类广泛分布于组织细胞内的天然低分子多价生物活性物质,在动物的消化、营养代谢以及正常生长中发挥重要作用。本实验克隆了猪(Sus scrofa)的7个与多胺代谢相关的基因,分别是精氨酸脱羧酶(ADC)、鸟氨酸脱羧酶(ODC)、鸟氨酸转氨酶(OAT)、鸟氨酸脱羧酶抗酶蛋白1~3(OAZ1、OAZ2、OAZ3)及多按调控因子-1(PMF1),基因片段长度分别是1 602(GenBank No. EU545649)、1 422(GenBank No. EU534186)、 1 465(GenBank No. EU404089)、 874(GenBank No. EU545195)、 678(GenBank No. EU545196)、667(GenBank No. EU545197)及703 bp(GenBank No. EU534185),开放阅读框的长度分别是1 380、1 383、1 313、681、568、559和615 bp,编码的氨基酸数目分别是460、461、439、227、189、186和205。经过比对分析,猪ADC基因编码的氨基酸序列与人、小鼠的同源性分别为89.7%和87%;猪ODC与人(Homo sapiens)、小鼠(Mus musculus)和大鼠(Rattus norvegicus)的同源性分别为93.1%、89.8%和90.5%;猪OAT与人、小鼠的同源性分别为91.8%、90.5%和89.8%;猪OAZ1与人、大鼠及小鼠的同源性分别为93.4%、82.5%和83.8%;猪OAZ2与人、大鼠及小鼠的同源性分别为99.5%、98.9%和98.9%;猪OAZ3与人、小鼠和大鼠的同源性分别为90.4%、87.5%和84.2%;猪PMF1基因编码的氨基酸序列与人和小鼠的同源性为84%和70.9%。半定量RT-PCR分析表明,断乳后ADC mRNA的表达量在盲肠和直肠有所增加,其他各组织均有所降低;ODC mRNA的表达量在直肠和盲肠有较大增加,结肠、回肠和空肠的表达量均呈小幅降低;OAT mRNA在肾的表达量有明显增加,十二指肠、空肠和回肠都有降低;除了胃,OAZ1 mRNA在其他组织表达水平均有所增加;OAZ2 mRNA的表达量在所有组织均有不同程度增加,盲肠居首;OAZ3mRNA在肠道组织的表达量均有不同程度增加;PMF1 mRNA的表达量在结肠、空肠和回肠与抗酶家族相似,但断乳后有小幅降低。研究结果提示,多胺对仔猪的生长、发育、成熟、适应性和损伤后的恢复具有重要的生物学作用
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郭豫杰
张志强
王月影
李宏基
查光明
李奎
杨国宇
关键词 仔猪多胺断乳克隆表达    
Abstract:Polyamines are ubiquitous organic cations of low molecular weight. The role of polyamines in the animal digestion, nutrient metabolism and normal growth is considered. We cloned polyamine metabolism-related genes polyamine-modulated factor 1(PMF1), arginine decarboxylase(ADC), ornithine decarboxylase(ODC), ornithine aminotransferase(OAT), ornithine decarboxylase antizyme 1~3(OAZ1, OAZ2 and OAZ3), their fragment length were 1 602(GenBank No. EU545649), 1 422(GenBank No. EU534186), 1 465(GenBank No. EU404089), 874(GenBank No. EU545195), 678(GenBank No. EU545196), 667(GenBank No. EU545197) and 703 bp(GenBank No. EU534185), respectively, with the open reading frame(ORF) of 1 380, 1 383, 1 313, 681, 568, 559 and 615 bp and, coding amino acids of 460, 461, 439, 227, 189, 186 and 205, respectively. The results of comparative analysis illustrated that the porcine ADC encoding of amino acid sequence shared 89.7% and 87.4% similar to that of human (Homo sapiens) and mouse(Mus musculus) respectively; the porcine ODC one shared 93.1% , 89.8% and 90.5% similar to that of human, mouse and rat respectively; the porcine OAT one shared 91.8%, 90.5% and 89.8% similar to that of human, mouse and rat (Rattus norvegicus) respectively; the porcine OAZ1 one shared 92.5%, 85.1% and 84.3% similar to that of human, mouse and rat respectively; the porcine OAZ2 one shared 98.1%, 97.9% and 97.2% similar to that of human, mouse and rat respectively; the porcine OAZ3 one shared 90.4%, 87.5% and 84.2% similar to that of human, mouse and rat respectively; the porcine PMF1 one shared 84% and 70.9% similar to that of human and mouse respectively. The analysis of the semi-quantitative RT-PCR showed that, after weaning, ADC mRNA increased significantly in appendix and rectal, while decreased slightly in other tissues; ODC mRNA increased significantly in appendix and rectal, while decreased slightly in jejunum, ileum and colon; OAT mRNA increased significantly in renal, while decreased slightly in duodenum, jejunum and ileum; OAZ1 mRNA increased in different magnitude, except in stomach; OAZ2 mRNA increased in different magnitude, especially in the appendix; OAZ3 mRNA increased in different magnitude in the intestinal tissues; PMF1 mRNA was similar to the (AZ) family in jejunum, ileum and colonic, but decreased slightly. These results suggest that polyamine are involved in the effect of the growth, development, mature, adaptability and repair of injured in piglet
Key wordsPiglet    Polyamine    Weaning    Cloning    Expression
收稿日期: 2012-03-27     
通讯作者: 杨国宇   
引用本文:   
郭豫杰1,张志强1,王月影1,李宏基1,查光明1,李奎 2,杨国宇1. 仔猪多胺代谢相关基因的克隆及断奶对其mRNA表达的影响[J]. , 2012, 20(8): 893-900.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I8/893
 
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