Cloning, Sequencing and Prokaryotic Expression of Homo Sapiens Heat Shock 10 kDa Protein and Mus Heat Shock 10 kDa Protein

doi:10.7668/hbnxb.2011.03.012

Abstract

Abstract: RT-PCR was used to study the expression of heat-shock 10 kDa protein.The results showed that there were two different HSP transcriptons of varied sizes in Homo liver cancer SMMC-7721 and Mus NS0.One HSP cDNA was composed of 312 bp(named HSPE1-1) and held 99% homology with HSPE1 mRNA reported(NM-002157).The other HSP gene cDNA was consisted of 313 bp(named Hspe1-1) and had 97% homology with Hspe1 mRNA reported(NM-008303).The homology between HSPE1-1 and Hspe1-1 was 92% in nucleotide sequence and 97% in amino acid sequence.Prokaryotic expression plasmids pET-28a-HSPE1-1 and pET-28a-Hspe1-1 for producing HSPE1 and Hspe1 proteins were constructed successfully.Its molecular weight was 14 kDa.Anti-His6 monoclonal antibody was used to demonstrate the identity of the proteins in Western Blot.The corresponding proteins were obtained after purification by Ni-NTA chromatography.The purified corresponding proteins would be helpfulin further studies of their molecular structure,biological function and clinical application.

Key words: Heat-shock 10 kDa protein ( Hsp10) , Homo sapiens heat shock 10 kDa protein 1 ( chaperonin 10) ( HSPE1) , Mus musculus heat shock protein 1 ( chaperonin 10) ( Hspe1) , Gene cloning, Protein expression

CLC Number:

WANG Li, DU Xiao-ming, WANG Lin-lin, SHI Xi-bao, TENG Man, LI Gong-quan, QUAN Kai, GUO Jun-qing, ZHANG Gai-ping. Cloning, Sequencing and Prokaryotic Expression of Homo Sapiens Heat Shock 10 kDa Protein and Mus Heat Shock 10 kDa Protein[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(3): 56-59. doi: 10.7668/hbnxb.2011.03.012.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2011.03.012

http://www.hbnxb.net/EN/Y2011/V26/I3/56

References

[1] Lindquist J,Craig E A.The heat shock proteins[J].Ann Rev Genet,1988,22:631-637.

[2] Bech mann R P,Mizzim I A,Welch W J.Interaction of folding and assen by events[J].Science,1990,248:850-854.

[3] 权凯,张改平.早孕因子的生物学特性及其在动物繁殖领域中的应用[J].中国畜牧兽医,2009,36(4):122-124.

[4] 李清州,郝惠芳,王丽,等.H5N1亚型禽流感病毒血凝素(HA)基因的表达及纯化[J].河南农业科学,2008(8):127-130.

[5] 万博,鲍登克,乔松林,等.猪α干扰素的表达及在PAM细胞系中抗PRRSV活性检测[J].河南农业科学,2010(3):93-96.

[6] 王秋霞,张美英,张改平,等.猪囊尾蚴排泄分泌抗原Ts881蛋白原核表达条件的优化[J].华北农学报,2009,24(3):59-63.

[7] 程晓丽,神吉智丈,李春燕,等.热休克蛋白10和热休克蛋白的60表达、纯化及与线粒体DNA转录因子A相互结合[J].郑州大学学报,2007,42(1):50-52.

[8] Yamada T,Nakamura R,Kidok.Function of hsp90 in differentiation and apoptosis of human Ec cells[J].Trans Soc Pathol,1995,84 (1):217-221.

[9] Graner M,Raymond A,Romney D,et al.Immunoprotoctive activities of multiple chaperone proteins isolated from murine B-cellleukemia/lymphoma[J].Clin Cancer Res,2000,6(3):909.

[10] Simons C,Claudia C,Rita A,et al.Human Hsp10 and early pregnancy factor (EPF) and their relationship andinvolvement in cancer and immunity:Current knowledge and perspectives[J].Life Sciences,2010,86:145-152.

[11] Ramona Dickson,Barbara Larsen,Paul V,et al.Cloning,expression,and purificatioonf afunctional nonacetylated mammalian mitochondrial chaperonin 10[J].The Journal of Biological Chemistry,1994,269(43):26858-26864.

[12] 苏岭,李绍戊,王荻,等.半定量RT-PCR方法检测热应激对鲫鱼肝脏中HSP70 mRNA含量的影响[J].华北农学报,2010,25(S1):106-110.

[1]	MA Jun, LIAN Kaiqi, SHAN Jinyan, LIU Yuling, LI Xiaolong, ZHAO Liming, LI Xuejun, PENG Renhai. *Cloning and Expression Analysis of bmp6 Gene in Carassius auratus* var.Qihe** [J]. Acta Agriculturae Boreali-Sinica, 2025, 40(2): 218-229.
[2]	SIBINUR Yashengjiang, ZHE Yuqi, ZHONG Jincheng, WU Zhijuan, CHAI Zhixin. ELOVL3 Gene Cloning,Eukaryotic Expression Vector Construction and Tissue Expression Profiling Analysis in Jiulong Yak [J]. Acta Agriculturae Boreali-Sinica, 2025, 40(2): 201-209.
[3]	HUANG Youju, YU Yongbo, PANG Cuijing, SUN Shixu, LU Chen, YU Yanchong. Cloning and Biology Function Analysis of Soybean GmWRKY44 [J]. Acta Agriculturae Boreali-Sinica, 2025, 40(1): 29-35.
[4]	LI Junren, CHEN Xiuzhen, WU Daidi. Cloning,Subcellular Localization,and Expression Analysis of PatASIL2 Gene of Trihelix Family from Pogostemon cablin [J]. Acta Agriculturae Boreali-Sinica, 2025, 40(1): 104-112.
[5]	LIN Hongjun, LÜ Ke, PAN Pingping, DONG Lili, XU Zhihao, WANG Zhonghua. *Cloning and Expression of the Anoectochilus roxburghii C4H* Gene and Its Role in the Flavonoid Synthesis Pathway** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(S1): 31-38.
[6]	YU Daoning, WANG Tong, LOBSANG Dondrub, PINGCUO Zhandui, ZHANG Qiang, ZHUOMA Ciren, NIMA Jiacuo, ZHANG Derong, LIANG Chunnian. *Cloning,Bioinformatics and Tissue Expression Analysis of PID1* Gene in Yak** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(6): 216-223.
[7]	LI Junren, WU Daidi, ZHAO Gongzheng, CHEN Xiuzhen. Cloning,Subcellular Localization and Expression Analysis of PatGTL1 Gene of Trihelix Family from Pogostemon cablin [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(6): 84-93.
[8]	ZHAO Anqi, YIN Yue, HE Jun, AN Wei, QIN Xiaoya, HU Tixu. Cloning,Subcellular Localization and Expression Analysis of LbaHY5 Gene from Lycium barbarum [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(6): 76-83.
[9]	GONG Yihui, ZHANG Canmei, ZHOU Guihua, CHEN Meiyu, TAN Qian, PI Shuiqin, ZENG Yongxian. *Cloning,Subcellular Localization and Expression Analysis of PpLAC7* Gene from Jinxiu Yellow Peach** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(5): 72-79.
[10]	GAO Wanting, LIU Zhida, SUN Xuetao, LI Zhiping, LÜ Wenxia, LI Aizhen, ZHAO Jun, ZHANG Zhiwei. *Cloning and Expression Patterns of StRac5,StRac7* and StRac13 Genes in Potato** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(5): 33-41.
[11]	GAN Lu, XIE Meijuan, LU Zhenhua, LI Ming, DING Bo, QIU Lina, XIE Xiaodong, WANG Junbin. *Cloning and Stress-Resistance Functional Analysis of Calcium-Dependent Protein Kinase Gene TaCDPK17* in Wheat** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(5): 1-8.
[12]	WANG Guanglong, XU Wujun, CHEN Yangyang, HU Zhenzhu, SUN Min, XIONG Aisheng. *Isolation and Functional Analysis of Garlic Calmodulin Genes AsCML15* and AsCML42 under Osmotic Stress** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 80-86.
[13]	ZHANG Yuanchen, SU Shengying, ZHANG Jiaqi, XUE Shuang, WANG Jingshun. *Cloning and Expression Analysis of MseLYS* from Oriental Armyworm,Mythimna separata(Lepidotera:Noctuidae)** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 206-214.
[14]	ZHOU Lin, LI Tuojian, QU Yan. Cloning and Expression Analysis of MiUGT79B1 and MpUGT79B1 Genes from Two Varieties of Meconopsis [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 102-109.
[15]	WANG Haiju, CHEN Xiaojuan, LI Tuojian, LUO Jun, QU Yan. *Cloning and Expression Analysis of DFR Gene from Meconopsis* with Different Colors** [J]. Acta Agriculturae Boreali-Sinica, 2024, 39(3): 88-95.

Please choose a citation manager

Content to export