JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2018, Vol. 53 ›› Issue (11): 1-8.doi: 10.6040/j.issn.1671-9352.0.2018.200

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Expression, purification and crystallization of gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa

Bo-ying XU1,2,*(),Tian-tian ZHANG1   

  1. 1. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China
    2. School of Life Sciences, University of Science and Technology of China, Heifei 230026, Anhui, China
  • Received:2018-04-17 Online:2018-11-01 Published:2018-11-14
  • Contact: Bo-ying XU E-mail:xbywh@126.com
  • Supported by:
    重庆市前沿与应用基础研究计划资助项目(cstc2015jcyjBX0142);重庆市前沿与应用基础研究计划资助项目(cstc2016jcyjA0375)

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Abstract:

Preliminary crystallographic study on the gas vesicle structure protein GvpC from the cyanobacterium Microcystis aeruginosa was performed. Firstly, gene gvpC was cloned and expressed; then, the recombinant protein was purified by denaturation and renaturation and gel filtration chromatography; the result of renaturation was detected by circular dichroism. Crystal screening was performed by sitting drop vapor diffusion techniques. GvpC was overexpressed as inclusion body in E. coli, and then the protein has been purified through denaturation and renaturation. The result of circular dichroism indicated that GvpC was refolded successfully in vitro. Crystals of GvpC were also obtained by crystal screening. Hence, the three-dimensional structure of GvpC could be determinated by solving its crystal structure.

Key words: Microcystis aeruginosa, gas vesicle, GvpC, denaturation and renaturation, protein crystal

CLC Number: 

  • Q816

Fig.1

The primary structure of GvpC(A) and amino acid sequence alignment of GvpC (B)"

Fig.2

The secondary structure of GvpC predicted by PSIPREDv3.3"

Fig.3

The tertiary structure prediction model of GvpC 33RR"

Fig.4

Expression of gvpC in E. coli"

Fig.5

Profile of size-exclusive chromatography of GvpC (A)and SDS-PAGE analysis of GvpC after SEC (B)"

Fig.6

Profile of the CD result of GvpC"

Table 1

The CD result of GvpC"

Secondary elements Ratio/%
Helix 30.0
Beta 0.0
Turn 31.2
Random 38.8
Total 100.0
RMS 35.479

Fig.7

Crystals of GvpC"

Fig.8

Multiple-sequence alignment of GvpC proteins from heterocyst-forming cyanobacteria"

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[1] XU Bo-ying, ZHANG Tian-tian. Heterelogous expression, purification andcrystal growth of gas vesicle protein GvpW from the cyanobacterial Microcystis aeruginosa [J]. JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE), 2017, 52(9): 103-110.
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