小尾寒羊DNAJC28基因克
发布时间：2018-11-28 10:00
【摘要】：DNAJC28(DnaJhomolog,subfamilyC,member28)是热激蛋白(Hsp40)家族的一员,是热激蛋白(HSP70)的分子伴侣,因HSP40都含有特殊的J结构域,所以又称为DNAJ。根据分子量大小,可以将热激蛋白分为6个亚家族:Hsp100、Hsp90、Hsp70、Hsp60、Hsp40和小热休克蛋白(如HSP27)。其中Hsp40由于拥有J结构域又被称为DNAJ。本次研究中,我们克隆鉴定了Hsp40家族的一个新成员,DnaJ(Hsp40)homolog,subfamilyC,member28,又名DNAJC28。热激蛋白是动物抗应激反应机制的重要组成部分,对于绵羊抗应激过程起到很关键的作用。DNAJ最初发现于大肠埃希氏杆菌,在真核生物中称为Hsp40,是Hsp70亚类辅助因子,主要调节Hsp70酶的活性。DNAJC28基因是DNAJ家族的一个基因,对于调节相应的Hsp70蛋白生物学功能起到关键的作用。本研究采用RT-PCR、RACE、荧光定量PCR等方法克隆了DNAJC28基因的全长序列,并在mRNA水平和蛋白水平进行了分析,主要研究结果如下:(1)本研究采用RACE等方法,以小尾寒羊为实验材料,克隆了小尾寒羊DNAJC28基因的全长cDNA序列。该基因cDNA全长为2729bp,3’端1228bp,5’端358bp,cds1143bp,开放阅读框编码380个氨基酸。(2)检测到DNAJC28基因在小尾寒羊和杜泊羊中呈现出组织表达差异性。在相同品种不同组织中表达差异显著,检测到在主动脉、肺脏、肾脏表达量较高,而在背最长肌、股二头肌等组织中表达量较低。(3)DNAJC28氨基酸系统进化同源性分析显示,该蛋白在哺乳动物中高度保守,绵羊该基因与牛、山羊相近而与蝙蝠家鼠等相距较远。综上所述,本研究成功克隆了小尾寒羊DNAJC28基因,对该基因序列及其编码的氨基酸序列的结构特征进行了预测,并比较分析了小尾寒羊和杜泊羊DNAJC28基因mRNA水平和蛋白水平上的组织表达谱特征,为后续研究该基因的功能提供了一定的依据
[Abstract]:DNAJC28 (DnaJhomolog,subfamilyC,member28) is a member of the heat shock protein (Hsp40) family and a chaperone of the heat shock protein (HSP70). Because HSP40 contains a special J domain, it is also called DNAJ.. According to the molecular weight, heat shock proteins can be divided into six subfamilies: Hsp100,Hsp90,Hsp70,Hsp60,Hsp40 and small heat shock proteins (such as HSP27). Hsp40 is called DNAJ. because it has J domain. In this study, we cloned and identified a new member of the Hsp40 family, DnaJ (Hsp40) homolog,subfamilyC,member28, also known as DNAJC28. Heat shock protein (HSPs) is an important part of the mechanism of animal anti-stress response, and plays a key role in the anti-stress process of sheep. DNAJ was first found in Escherichia coli, and in eukaryotes, Hsp40, is a cofactor of Hsp70 subclass. DNAJC28 gene is a gene of DNAJ family and plays a key role in regulating the biological function of Hsp70 protein. In this study, the full-length sequence of DNAJC28 gene was cloned by RT-PCR,RACE, fluorescence quantitative PCR, and analyzed at mRNA level and protein level. The main results were as follows: (1) RACE was used in this study. The full-length cDNA sequence of DNAJC28 gene was cloned from small tail Han sheep. The total length of the cDNA is 2729bp1 228bpl5'end 358bpcds1143bp. the open reading frame encodes 380 amino acids. (2) the difference of tissue expression of DNAJC28 gene was detected in small tail Han sheep and du Bo sheep. The expression level was higher in aorta, lung and kidney, but lower in longissimus dorsi and biceps femoris. (3) phylogenetic homology analysis of DNAJC28 amino acid system. This protein is highly conserved in mammals. The gene of sheep is close to cattle and goat and is far away from bat and mouse. In conclusion, the DNAJC28 gene of small tail Han sheep was cloned successfully, and the structural characteristics of the gene and the amino acid sequence were predicted. The tissue expression profiles of DNAJC28 gene mRNA and protein in small tail Han sheep and du Bo sheep were compared and analyzed, which provided a basis for further study on the function of this gene.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S826
本文编号：2362523
[Abstract]:DNAJC28 (DnaJhomolog,subfamilyC,member28) is a member of the heat shock protein (Hsp40) family and a chaperone of the heat shock protein (HSP70). Because HSP40 contains a special J domain, it is also called DNAJ.. According to the molecular weight, heat shock proteins can be divided into six subfamilies: Hsp100,Hsp90,Hsp70,Hsp60,Hsp40 and small heat shock proteins (such as HSP27). Hsp40 is called DNAJ. because it has J domain. In this study, we cloned and identified a new member of the Hsp40 family, DnaJ (Hsp40) homolog,subfamilyC,member28, also known as DNAJC28. Heat shock protein (HSPs) is an important part of the mechanism of animal anti-stress response, and plays a key role in the anti-stress process of sheep. DNAJ was first found in Escherichia coli, and in eukaryotes, Hsp40, is a cofactor of Hsp70 subclass. DNAJC28 gene is a gene of DNAJ family and plays a key role in regulating the biological function of Hsp70 protein. In this study, the full-length sequence of DNAJC28 gene was cloned by RT-PCR,RACE, fluorescence quantitative PCR, and analyzed at mRNA level and protein level. The main results were as follows: (1) RACE was used in this study. The full-length cDNA sequence of DNAJC28 gene was cloned from small tail Han sheep. The total length of the cDNA is 2729bp1 228bpl5'end 358bpcds1143bp. the open reading frame encodes 380 amino acids. (2) the difference of tissue expression of DNAJC28 gene was detected in small tail Han sheep and du Bo sheep. The expression level was higher in aorta, lung and kidney, but lower in longissimus dorsi and biceps femoris. (3) phylogenetic homology analysis of DNAJC28 amino acid system. This protein is highly conserved in mammals. The gene of sheep is close to cattle and goat and is far away from bat and mouse. In conclusion, the DNAJC28 gene of small tail Han sheep was cloned successfully, and the structural characteristics of the gene and the amino acid sequence were predicted. The tissue expression profiles of DNAJC28 gene mRNA and protein in small tail Han sheep and du Bo sheep were compared and analyzed, which provided a basis for further study on the function of this gene.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S826
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