Panhandle PCR strategy to amplify the upstream unknown sequence of the Pr1 gene of pythium guiyangense

DUAN Siliang; SU Xiaoqing; YU Sheng

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Panhandle PCR strategy to amplify the upstream unknown sequence of the Pr1 gene of pythium guiyangense

Updated：2026-03-12

- Panhandle PCR strategy to amplify the upstream unknown sequence of the Pr1 gene of pythium guiyangense
- Military Medical Research Vol. 28, Issue 5, Pages: 268-272(2013)
- Affiliations：
  
  1. Medical College, Guangxi University of Science and Technology
  2. Department of Biology, Guiyang Medical College
- Author bio：
- Funds：
- DOI：
  CLC： Q933
- Published：2013
- Accepted：
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[1]DUAN Siliang,SU Xiaoqing,YU Sheng.Panhandle PCR strategy to amplify the upstream unknown sequence of the Pr1 gene of pythium guiyangense[J].Journal of Medical Colleges of PLA,2013,28(05):268-272.
DOI：

[1]DUAN Siliang,SU Xiaoqing,YU Sheng.Panhandle PCR strategy to amplify the upstream unknown sequence of the Pr1 gene of pythium guiyangense[J].Journal of Medical Colleges of PLA,2013,28(05):268-272. DOI：

摘要

Abstract

Objective: Based on a partialsubtilisin-like protease

Pr1 genomic sequence of Pythium guiyangense which has been cloned before

Panhandle PCR strategy was used to amplify the upstream flanking sequence adjacent to the known sequence of the Pr1gene. Methods: The genomic DNA was firstly digested with BamHⅠ and then treated with calf intestinal alkaline phosphatase（CIAP）. Next

a 5′ phosphorylated oligonucleotide was ligated to the 5′ ends of BamHⅠ-digested DNA. After denaturation

intrastrand annealing and polymerase extension

a pan with a handle was formed

and lastly the nested PCR was performed. Results: A 864 bp product was amplified

which was adjacent to the known sequence of Pr1 gene.The gene has been accessed by GenBank（Accession:JQ975036）. Conclusion: Panhandle PCR is a quick and convenient approach for amplifying and identifying unknown partner genes

which facilitates cloning full-length Pr1 gene.

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