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Construction of Prokaryotic Expression Vector Conjugatedwith B56α Regulatory Subunit of PP2A(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2016年03期
Page:
74-
Research Field:
·生命科学·
Publishing date:

Info

Title:
Construction of Prokaryotic Expression Vector Conjugatedwith B56α Regulatory Subunit of PP2A
Author(s):
Zhao Yaping; Cheng Xiaoqing; Sun Xiaoli; Li Liangyuan; Zhang Zhao
School of Life Sciences，Nanjing Normal University，Jiangsu Key Laboratory for Molecular and Medical Biotechnology，Nanjing 210023，China
Keywords:
PP2A; B56α; prokaryotic expression
PACS:
Q28
DOI:
10.3969/j.issn.1001-4616.2016.03.013
Abstract:
pCEP-4HA-B56α was used as a template to construct the prokaryotic expression vector of PP2A B56α，primers were designed according to cDNA sequence to clone the gene. The amplified cDNA fragment was inserted into pGEX-4T-1 vector. Positive clones were verified via sequencing. The recombinant plasmid was transformed into BL21 E.coli. Fragmentation of IPTG-induced E.coli culture was achieved by sonication on ice. Soluble and insoluble fractions were separated and analyzed. Lastly，soluble fraction was purified. Results showed that，the constructed plasmid contained the fragment of PP2A B56α. The recombinant protein was about 79 kD and soluble recombinant protein was about 8.6% of total bacteria protein. The purification of GST-tagged B56α was performed by use of GST purification system. The purity of recombinant protein was reached to 78.9% among total protein by using this method and the rate of recovery was 52.2%.The PP2A B56α Prokaryotic Expression System was constructed successfully and recombinant protein was obtained，which was helpful for the future study of its biological function.

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Last Update: 2016-09-30