小尾寒羊DNAJC28基因克
發(fā)布時間：2018-11-28 10:00
【摘要】：DNAJC28(DnaJhomolog,subfamilyC,member28)是熱激蛋白(Hsp40)家族的一員,是熱激蛋白(HSP70)的分子伴侶,因HSP40都含有特殊的J結(jié)構(gòu)域,所以又稱為DNAJ。根據(jù)分子量大小,可以將熱激蛋白分為6個亞家族:Hsp100、Hsp90、Hsp70、Hsp60、Hsp40和小熱休克蛋白(如HSP27)。其中Hsp40由于擁有J結(jié)構(gòu)域又被稱為DNAJ。本次研究中,我們克隆鑒定了Hsp40家族的一個新成員,DnaJ(Hsp40)homolog,subfamilyC,member28,又名DNAJC28。熱激蛋白是動物抗應(yīng)激反應(yīng)機制的重要組成部分,對于綿羊抗應(yīng)激過程起到很關(guān)鍵的作用。DNAJ最初發(fā)現(xiàn)于大腸埃希氏桿菌,在真核生物中稱為Hsp40,是Hsp70亞類輔助因子,主要調(diào)節(jié)Hsp70酶的活性。DNAJC28基因是DNAJ家族的一個基因,對于調(diào)節(jié)相應(yīng)的Hsp70蛋白生物學(xué)功能起到關(guān)鍵的作用。本研究采用RT-PCR、RACE、熒光定量PCR等方法克隆了DNAJC28基因的全長序列,并在mRNA水平和蛋白水平進(jìn)行了分析,主要研究結(jié)果如下:(1)本研究采用RACE等方法,以小尾寒羊為實驗材料,克隆了小尾寒羊DNAJC28基因的全長cDNA序列。該基因cDNA全長為2729bp,3’端1228bp,5’端358bp,cds1143bp,開放閱讀框編碼380個氨基酸。(2)檢測到DNAJC28基因在小尾寒羊和杜泊羊中呈現(xiàn)出組織表達(dá)差異性。在相同品種不同組織中表達(dá)差異顯著,檢測到在主動脈、肺臟、腎臟表達(dá)量較高,而在背最長肌、股二頭肌等組織中表達(dá)量較低。(3)DNAJC28氨基酸系統(tǒng)進(jìn)化同源性分析顯示,該蛋白在哺乳動物中高度保守,綿羊該基因與牛、山羊相近而與蝙蝠家鼠等相距較遠(yuǎn)。綜上所述,本研究成功克隆了小尾寒羊DNAJC28基因,對該基因序列及其編碼的氨基酸序列的結(jié)構(gòu)特征進(jìn)行了預(yù)測,并比較分析了小尾寒羊和杜泊羊DNAJC28基因mRNA水平和蛋白水平上的組織表達(dá)譜特征,為后續(xù)研究該基因的功能提供了一定的依據(jù)
[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.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S826
本文編號：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.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S826
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