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pYEMF, a pUC18-Derived XcmI T-Vector for Efficient Cloning of PCR Products

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Abstract

A 1330-bp DNA sequence with two XcmI cassettes was inserted into pUC18 to construct an efficient XcmI T-vector parent plasmid, pYEMF. The large size of the inserted DNA fragment improved T-vector cleavage efficiency, and guaranteed good separation of the molecular components after restriction digestion. The pYEMF-T-vector generated from parent plasmid pYEMF permits blue/white colony screening; cloning efficiency analysis showed that most white colonies (>75%) were putative transformants which carried the cloning product. The sequence analysis and design approach presented here will facilitate applications in the fields of molecular biology and genetic engineering.

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Abbreviations

MCS:

Multi-cloning site

IPTG:

β-d-1-Thiogalactopyranoside

X-Gal:

5-Bromo-4-chloro-3-indolyl-β-d-galactopyranoside

ORF:

Open reading frame

US:

Upstream sites

DS:

Downstream sites

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Acknowledgments

This study was supported by a General Program Grant from the National Natural Science Foundation of China (Grant No. 31070046), and the Science and Technology Research Grant Programs of University of Jinan (Grant Nos. B0301, B0524).

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Authors and Affiliations

  1. Biotechnology Department, School of Medicine and Life Sciences, University of Jinan, 106 Jiwei Road, Jinan, 250022, China

    Jingsong Gu & Chunjiang Ye

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  1. Jingsong Gu
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  2. Chunjiang Ye
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Correspondence to Chunjiang Ye.

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Gu, J., Ye, C. pYEMF, a pUC18-Derived XcmI T-Vector for Efficient Cloning of PCR Products. Mol Biotechnol 47, 229–233 (2011). https://doi.org/10.1007/s12033-010-9333-y

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