期刊文献+

杆状病毒蛋白表达系统制备可溶性LRIG1蛋白及其鉴定 被引量:3

Preparation of soluble leucine-rich repeats and immunoglobulin-like domains 1 protein via the Baculovirus expression vector system and its identification
下载PDF
导出
摘要 目的为进一步研究肿瘤的治疗提供高质量的可溶性多亮氨酸重复区免疫球蛋白样蛋白(sLRIG1)。方法用重组sLRIG1的杆状病毒(Bac-sLRIG1)感染sf9昆虫细胞,表达的蛋白经提纯,获得sLRIG1蛋白,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹进行分析鉴定。结果使用杆状病毒表达载体系统(BEVS)成功表达了重组sLRIG1蛋白,最佳感染复率为5,孵育时间为72 h,经鉴定蛋白表达正确,纯度高达95%。结论用BEVS可成功获得大量高纯度重组sLRIG1蛋白,可满足后续实验的需要。 Objective To provide high quality soluble leucine-rich repeats and immunoglobulin-like domains 1 (sLRIG1) protein for further research on the treatment of malignant tumors. Methods Sf9 insect cells were infected with the recombinant sLRIG1 Baculovirus encoding the extra-cellular domain of human LRIG1 (gene No. AF381545). The supernatant of the culture medium was collected and the recombinant sLRIG1 was purified by the Ni Sepharose Fast Flow Column and the DEAE Sepharose Fast Flow Column. The obtained protein was identified by SDS-PAGE technique and Western blotting. Results The sLRIG1 protein was successfully expressed and its expression reached peak at 72 hours after infection when the multiplicity of infection was 5. The purity was over 95%. Conclusion The sLRIG1 protein was successfully preparated via Baculovirus expression vector system. The purified protein had a high quality and was suitable for the further research on the treatment of malignant tumors.
出处 《中国临床神经外科杂志》 2012年第8期476-478,共3页 Chinese Journal of Clinical Neurosurgery
基金 教育部博士点新教师基金No.200804861039 国家自然科学基金No.30973073
关键词 可溶性LRIG1 杆状病毒蛋白表达系统 蛋白表达 纯化 鉴定 Soluble leucine-rich repeats and immunoglobulin-like domains 1 (sLRIG1) Baculovirns Expression Vector System (BEVS) Preparation Identification
  • 相关文献

参考文献11

  • 1Suzuki Y, Sato N, Tohyama M, et al. cDNA cloning of a novel membrane glycoprotein that is expressed specifically in glial cells in the mouse brain. LIG-1, a protein with leu- cine-rich repeats and immunoglobulin-like domains [J]. J Biol Chem, 1996, 271: 22522-22527.
  • 2Nilsson J, Vallbo C, Guo D, et al. Cloning, characterization, and expression of human LIGI [J]. Biochem Biophys Res Commun, 2001, 284:1155-1161.
  • 3Guo D, Hohnlund C, Henriksson R, et al. The LRIG gene family has three vertebrate paralogs widely expressed in human and mouse tissues, and a homolog in Ascidiacea [J]. Genomics, 2004, 84: 157-165.
  • 4Yi W, Ye F, Guo DS, et al. Expression of leucine-rich re- peats and immunoglobulin-like domains (LRIG) proteins in human astrocytomas [J]. Chinese-German J Clin Oncol, 2005, 4(4): 225-228.
  • 5Hedman H, Henriksson R. LRIG inhibitors of growth factor signalling - double-edged swords in human cancer [J]. Eur J Cancer, 2007, 43(4): 676-682,.
  • 6Hedman H, Nilsson J, Guo D, et al. Is LRIGI a tumour sup- pressor gene at chromosome 3p14.3 [J]? Acta Oncol, 2002, 41 (4): 352-354.
  • 7易伟,叶飞,郭东升,薛德麟,雷霆.LRIG1基因在人星形细胞瘤中的表达与意义[J].中国临床神经外科杂志,2006,11(1):29-31. 被引量:7
  • 8Yi W, Holmlund C, Nilsson J, et al. Paracrine regulation of growth factor signaling by shed leucine-rich repeats and immunoglobulin-like domains 1 [J]. Exp Cell Res, 2011,317(4): 504-512.
  • 9Goldoni S, Iozzo RA, Kay P, et al. A soluble ectodomain of LRIG1 inhibits cancer cell growth by attenuating basal and ligand-dependent EGFR activity [J]. Oncogene, 2007, 26 (3): 368-381.
  • 10O'Reilly D, Miller LK, Luckow VA. Baculovirus Expression Vectors: A Laboratory Manual [M]. New York: Freeman and Company, 1992.32-36.

二级参考文献13

  • 1Nilsson J, Vallbo C, Guo D, et al . Cloning, characterization, and expression of human LIGI [J]. Biochem Biophys Res Commun, 2001, 284(5): 1155-1161.
  • 2Hedman H, Nilsson J, Guo D, Henriksson R. Is LRIG1 a tumour suppressor gene at chromosome 3p14.3 [J]? Acta Oncol, 2002, 41(4): 352-354.
  • 3Kleihues P, Louis DN, Scheithauer BW, et al . The WHO classification of tumors of the nervous system [J]. J Neuropathol Exp Neurol, 2002, 61(3): 215-229.
  • 4Musacchio M, Perrimon N. The Drosophila kekkon genes:novel members of both the leucine-rich repeat and immunoglobulin superfamilies expressed in the CNS. Dev Biol,1996, 178(1): 63-76.
  • 5Suzuki Y, Sato N, Tohyama M, et al . cDNA cloning of a novel membrane glycoprotein that is expressed specifically in glial cells in the mouse brain. LIG-1, a protein with leucine-rich repeats and immunoglobulin-like domains [J].J Biol Chem, 1996, 271(37): 22522-22527.
  • 6Maga G, Hubscber U. Proliferating cell nuclear antigen (PCNA): a dancer with many partners [J]. J Cell Sci, 2003, 116(Pt15): 3051-3060.
  • 7Ghiglione C, Canaway KL 3rd, Amundadottir LT, et al, .The transmembrane molecule kekkon 1 acts in a feedback loop to negatively regulate the activity of the Drosophila EGF receptor during oogenesis [J]. Cell, 1999, 96(6): 847-856.
  • 8Mukohara T, Kudoh S, Yamauchi S, et al . Expression of epidermal growth factor receptor (EGFR) and downstream-activated peptides in surgically excised non-small-cell lung cancer (NSCLC) [J]. Lung Cancer, 2003, 41(2): 123-130.
  • 9Mimeault M, Pommery N, Henichart JP. New advances on prostate carcinogenesis and therapies: involvement of EGF-EGFR transduction system [J]. Growth Factors, 2003,21(1): 1-14.
  • 10Tsutsui S, Ohno S, Murakami S, et al . Comparison of the immunohistochemical expression of EGFR, c-erbB2 and p53 protein between primary and recurrent breast cancer[J]. Breast Cancer, 2002, 9(2): 111-117.

共引文献6

同被引文献22

  • 1易伟,叶飞,郭东升,薛德麟,雷霆.LRIG1基因在人星形细胞瘤中的表达与意义[J].中国临床神经外科杂志,2006,11(1):29-31. 被引量:7
  • 2Gur G, Rubin C, Katz M, et al. LRIG1 restricts growth factor signaling by enhancing receptor ubiquitylation and degra- dation [J]. EMBO J, 2004, 23(16): 3270-3281.
  • 3Shattuck DE, MilLer .IK, Laederich M, et al. LRIG1 is a novel negative regulator of the Met receptor and opposes Met andHer2 synergy [J]. Mol Cell Biol, 2007, 27(5): 1934-1946.
  • 4Ledda F, Bieraugel O, Fard SS, et al. LRIG1 is an endoge- nous inhibitor of Ret receptor tyrosine kinase activation, downstream signaling, and biological responses to GDNF [J]. J Neurosci, 2008, 28(1): 39-49.
  • 5Nilsson J, Vallbo C, Guo D, et al. Cloning, characterization, and expression of human LR1G1 [J]. Biochem Biophys Res Commun, 2001, 284:1155-1161.
  • 6Sathornsumetee S, Reardon DA, Desjardins A, et al. Mole- cularly targeted therapy for malignant glioma [J]. Cancer, 2007, 110(1): 13-24.
  • 7Omuro AM, Faivre S, Raymond E. Lessons learned in the development of targeted therapy for malignant gliomas [J]. Mol Cancer Ther, 2007, 6(7): 1909-1919.
  • 8Yi W, Ye F, Guo D, et al. Expression of leucine-rich repeats and immunoglobulin-like domains(LRIG) proteins in human astrocytomas [J]. Chin-Ger J Clin Oncol, 2005, 4(4): 225- 228.
  • 9Hedman H, Henriksson R. LRIG inhibitors of growth factor signalling- double-edged swords in human cancer [J]? Eur J Cancer, 2007, 43(4): 676-682.
  • 10Hedman H, Nilsson J, Guo D, et al. Is LRIG1 a tumour sup- pressor gene at chromosome 3p14.3 [J]? Acta Oneol, 2002, 41(4): 352-354.

引证文献3

二级引证文献3

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部