New insight into the method of posttransformational vector amplification (PTVA) in Pichia pastoris

Liangcheng Jiao; Qinghua Zhou; Wu Liu; Yunjun Yan

doi:10.1016/j.mimet.2018.04.013

New insight into the method of posttransformational vector amplification (PTVA) in Pichia pastoris

J Microbiol Methods. 2018 May:148:151-154. doi: 10.1016/j.mimet.2018.04.013. Epub 2018 Apr 17.

Authors

Liangcheng Jiao¹, Qinghua Zhou¹, Wu Liu¹, Yunjun Yan²

Affiliations

¹ Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
² Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China. Electronic address: [email protected].

PMID: 29678499
DOI: 10.1016/j.mimet.2018.04.013

Abstract

Posttransformational vector amplification (PTVA) is widely used to enrich the gene-copy number in Pichia pastoris. We engineered two test strains for PTVA studies and demonstrate that the PTVA process results in the amplification of a fragment with the resistant gene flanked by two homologous arms instead of the entire vector.

Keywords: Homologous arms; Homologous recombination; Multi-copy strains; Pichia pastoris; Posttransformational vector amplification.

MeSH terms

DNA Replication
Gene Dosage*
Genetic Vectors*
Pichia / genetics*
Transformation, Genetic*