Prokaryotic expression, purification, and identification of GST-β-catenin-His double labeled fusion protein

doi:10.7523/j.issn.2095-6134.2012.6.019

›› 2012, Vol. ›› Issue (6): 847-852.DOI: 10.7523/j.issn.2095-6134.2012.6.019

Prokaryotic expression, purification, and identification of GST-β-catenin-His double labeled fusion protein

YIN Hui-Long, YUAN Li

College of Life Sciences, Graduate University, Chinese Academy of Sciences, Beijing 100049, China

Received:2011-11-10 Revised:2012-01-13 Online:2012-11-15

Abstract

Abstract: The cDNA sequence of β-catenin was amplified through RT-PCR with 6xHis tag added in the gene downstream, cloned into the vector pGEX-4T-1. The plasmid transformed into BL21pLysS was induced by IPTG. The induced fusion protein was purified stepwise using Glutathione Sepharose 4B and Ni columns. The final protein ran on SDS-PAGE with a specific band at the expected size of 114 kDa. Western Blotting showed that the purified protein can be recognized by the anti-β-catenin, anti-GST, and anti-His antibodies, respectively. All these results provide a basis for further study of β-catenin function.

Key words: β-catenin, purification, GST and His tags, Western Blotting

CLC Number:

R346

YIN Hui-Long, YUAN Li. Prokaryotic expression, purification, and identification of GST-β-catenin-His double labeled fusion protein[J]. , 2012, (6): 847-852.

References

[1] Wang F, Li F, Yang A Q, et al. β-catenin and E-cadherin the expression and significance in ovarian serous tumor[J]. Journal of Nongken Medicine, 2011, 33(1):1-4(in Chinese). 王芳,李锋,杨安强,等. β-catenin和E-cadherin在卵巢浆液性肿瘤中的表达及意义[J]. 农垦医学,2011,33(1):1-4.
[2] Kajiyama H,Shibata K,Ino K,et al.Possible involvement of SDF lalpha/CXCR4-DPPIV axis in TGF-betal-induced enhancement of migratory potential in human peritoneal mesothelial cells[J].Cell Tissue Res,2007,330(2):221-229.
[3] Lin Y C,Wu M Y,Li D R,et al.Prognostic and elinicopadthological features of E-cadherin,α-catenin,β-catenin,γ-catenin and cycIin D1 expression in humanesophageal squamous cell carcinoma World[J]. Gastroenterol,2004,10(22):3235-3239.
[4] Polakis P.Wnt signaling and cancer[J]. Genes Dev,2000,14(15):1837-1851.
[5] Li H L, He Y J, Qiao C, et al. Expression and significance of β-catenin, cyclinD1 in breast carcinoma[J]. Shanxi Medical Journal, 2011, 40(6):558-560(in Chinese). 李洪利,何艳姣,乔超,等. 乳腺癌β-catenin cyclinD1的表达及意义[J]. 山西医药杂志,2011,40(6):558-560.
[6] Fieke E M,Christine F,Claude C,et al. Retinoic acid-induced pancreatic stellate cell quiescence reduces paracrine Wnt-β-catenin signaling to slow tumor progression[J]. Gastroenterology, 2011,141(4):1486-1497.
[7] Andrzej W,Mariusz K,Stanislaw S,et al. Comparison of Beta-catenin with TGF-beta1,HIF-1alpha and patients disease-free survival in Human colorectal cancer[J].Pathology & Oncology Research,2010,16(3):311-318.
[8] Zhao X L, Duester G. Effect of retinoic acid signaling on Wnt/β-catenin and FGF signaling during body axis extension[J]. Gene Expression Patterns,2009,9(6):430-435.
[9] Lindsay J G, Rob B. Redistribution of β-catenin in response to EGF and lithium signalling in human oesophageal squamous carcinoma cell lines[J]. Cancer Cell International,2003,3(1):13.

Prokaryotic expression, purification, and identification of GST-β-catenin-His double labeled fusion protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 5

Recommended Articles

Metrics

Comments

[1]	ZHANG Zheng, WANG Junfeng, MO Xiaojian, HU Weiwei. Experimental study of dust particulate collection by charged droplets [J]. , 2016, 33(2): 265-270.
[2]	DU Kun, SONG Qiucheng, QIAO Congfeng. Synchronous concentration and purification schemes of arbitrary unknown hyperentangled mixed states [J]. , 2015, 32(2): 158-165.
[3]	XU Feng, MU Xin, WANG Wei, LIU Li-Xin. Screening proteins interacting with hCLP46 using tandem affinity purification [J]. , 2011, 28(2): 210-216.
[4]	WANG Yun, ZENG Wo-Tan. Purification and characterization of β-glucosidase from Aspergillus niger [J]. , 2009, 26(2): 274-279.
[5]	QI Chao. Purification and Characterization of UGPase [J]. , 2004, 21(3): 345-351.