PMID- 18237532
OWN - NLM
STAT- MEDLINE
DCOM- 20100402
LR  - 20201209
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 24
IP  - 2
DP  - 2008 Feb
TI  - [Fusion expression of human beta defensin 4 in Escherichia coli and preparation 
      of its polyclonal antibody].
PG  - 150-2
AB  - AIM: To construct the prokaryotic fusion expression vector of human beta defensin 
      4 (HBD4), and to express GST-HBD4 fusion protein in Escherichia coli (E.coli) and 
      prepare polyclonal antibody of GST-HBD4. METHODS: The gene encoding mature 
      peptide of HBD4 (mHBD4) was amplified by PCR from cloning vector PMD18-T/HBD4 
      which contained the full-length HBD4 cDNA and then cloned into prokaryotic 
      expression vector PGEX-4T-2 to construct PGEX-4T-2/mHBD4. GST-HBD4 expression was 
      induced by IPTG. The antiserum was prepared by immunizing rabbit with 
      GST-HBD4.The titer and specificity of the antibody were detected by ELISA and 
      Western blot, respectively. RESULTS: The recombinant expression vector 
      PGEX-4T-2/mHBD4 was successfully constructed. After being induced by IPTG, The 
      fusion protein with relative molecular mass of 32 000 was successfully expressed 
      in E.coli and partly expressed in the soluble form in supernatant. The rabbit 
      antibody against GST-HBD4 was obtained. The ELISA titer of antiserum against 
      GST-HBD4 was about 1:128 000. Western blot analysis showed that the antiserum 
      could bind to the expressed GST-HBD4 specifically. CONCLUSION: The rabbit 
      antibody against GST-HBD4 has been successfully prepared, which lays the 
      foundation for further studying the structure and function of HBD4.
FAU - Cao, Yu-hong
AU  - Cao YH
AD  - Department of Pediatrics, Xijing Hospital, Fourth Military Medical University, 
      Xi'an 710032, China. caoyh70@126.com
FAU - Zhang, Guang-yun
AU  - Zhang GY
FAU - Zhang, Guo-cheng
AU  - Zhang GC
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Antibodies)
RN  - 0 (DEFB4A protein, human)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (beta-Defensins)
SB  - IM
MH  - Animals
MH  - Antibodies/*immunology/*metabolism
MH  - Blotting, Western
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Escherichia coli/genetics/*metabolism
MH  - Humans
MH  - Polymerase Chain Reaction
MH  - Rabbits
MH  - Recombinant Fusion Proteins/genetics/*immunology/*metabolism
MH  - beta-Defensins/genetics/*immunology/*metabolism
EDAT- 2008/02/02 09:00
MHDA- 2010/04/03 06:00
CRDT- 2008/02/02 09:00
PHST- 2008/02/02 09:00 [pubmed]
PHST- 2010/04/03 06:00 [medline]
PHST- 2008/02/02 09:00 [entrez]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Feb;24(2):150-2.