[Construction and identification of the recombinant prokaryotic expression plasmid containingbetapep-25 peptide].
OBJECTIVE
To synthesize antiangiogenic peptide fragment of betapep-25, to construct and identify the recombinant prokaryotic expression plasmid containing betapep-25 peptide.


METHODS
The fragment encoding betapep-25 peptide was designed and synthesized artificially and was cloned into vector pGEM-T easy after being identified by sequencing. After being digested by Nae I and BamH I, T-betapep-25 peptide fragment was cloned into recombinant vector pBV220-NT4, which was digested by Nae I and BamH I. The constructed recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 was digested using BamH I, EcoR I and BamH I, respectively.


RESULTS
The sequcence of betapep-25 synthesized artificially and analyzed by digestion was consistent with the published results. Fragments of 4,030 bp, 364 bp and 3,666 bp were obtained after digestion of recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 using BamH I, EcoR I and BamH I, respectively, which demonstrated the successful cloning and construction of recombinant prokaryotic expression plasmid.


CONCLUSIONS
The successful construction recombinant prokaryotic expression plasmid expressing betapep-25 provides a foundation for further research on its mechanism of anti-tumor activity in vivo and in vitro.
Li
En-xiao
EX
Department of Medical Oncology of the First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. lienxiao@csco.org.cn
Liu
Shan-xi
SX
Yang
Guang-xiao
GX
Wang
Quan-ying
QY
Wu
Yin-ying
YY
Shi
Fan
F
D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D011387:Prokaryotic Cells / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques
D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; C111993:betapep-25 protein, synthetic