title: pYEMF, a pUC18-derived XcmI T-vector for efficient cloning of PCR products.
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.
authors.affiliation: Biotechnology Department, School of Medicine and Life Sciences, University of Jinan, 106 Jiwei Road, Jinan 250022, China.
authors.forename: Jingsong
authors.initials: J
authors.lastname: Gu
authors.forename: Chunjiang
authors.initials: C
authors.lastname: Ye
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: C084267:endodeoxyribonuclease XcmI; D015252:Deoxyribonucleases, Type II Site-Specific