JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2009, Vol. 47 ›› Issue (6): 15-19.

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onstruction and identification of the expression vector ofNT4-SAC-HA2-TAT fusion gene

 ZHANG Shi-Bao, LIU Qing-Yong, RUAN Xi-Yun, CHEN Jie, ZHANG Jian-Jun, LI Zong-Wu, YANG Guang-Xiao, WANG Quan-Ying   

  1. 1. Department of Urinary Surgery, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China; 2. Department of Neurology, Jinan Central Hospital Affiliated to ShandongUniversity, Jinan 250013, China; 3. Xi'an Huaguang Biological Engineering Co.Ltd, Xi'an 710025, China
  • Received:2008-12-04 Published:2009-06-16

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Abstract:

Objective To construct the prokaryocyte expression vector bearing fusion gene NT4-SAC-HA2-TATand lay a foundation for further study on the genetic therapy of prostate cancer.  Methods  By means of asymmetrical primer/template, the fragment encoding SAC was gained by the PCR method. Then the SAC/NaeI, KpnI and HA2-TAT/KpnI, and Xho I were cloned into recombinant vector pBV220-NT4/NaeI,SalI and the recombinant plasmid pBV220-NT4-SAC-HA2-TATwas obtained. The fragment NT4-SAC-HA2-TATobtained was amplified by PCR. Then the fragment was easily cloned into vector pGEM-T and was sequenced by the dideoxy-mediated chain-termination method. The cloned NT4-SAC-HA2-TATcDNA was compared with the GeneBank sequence by the DNASIS.  Results By DNA sequencing, the sequence of SAC was obtained which included NaeI andKpnI restriction enzyme sites by pcr. The fusion gene NT4-SAC-HA2-TATwas successfully sub-cloned into pBV220.  Conclusion   The recombinant vector pBV220-NT4-SAC-HA2-TATwas successfully constructed. These results lay the foundation for further research of using fusion genes to treat prostate cancer.

Key words: Par-4; fusion peptide; Prostate neoplasms; Gene therapy

CLC Number: 

  • R737.25
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