Construction of human lung cancer metastasis related protein 1 recombinant lentiviral vector and virus packaging and identification
LI Yan-qin
YU Ling
LIANG Yuan
YU Qing-yang
LIU Yang
LI Chun-sun
Abstract：Objective To construct a lentiviral expressing vector carrying human lung cancer metastasis related protein 1 (LCMR1) gene and then the overexpression in lung cancer 95C cell line was tested in order to provide an experimental foundation for the function of LCMR1 on lung cancer.Methods LCMR1 gene cDNA was amplicated by polymerase chain reaction (PCR) and cloned into the pLVX-IRES-Neo vector.Reconstructed pLVX-IRES-Neo-LCMR1 plasmid was identified by electromigratory analysis,colony PCR and sequencing analysis.The viral stock was prepared by cotransfection of plasmids and the packaging plasmid mix to 293T cells.After infection of 95C cells with these lentiviruses,the expression of LCMR1 was detected by q-PCR and Western blot.Results LCMR1 gene was constructed into the pLVX-IRES-Neo vector successfully.Enzyme digestion got correct length of LCMR1 gene,DNA sequencing analysis confirmed that LCMR1 gene sequence was exactly the same with that reported by GenBank.There were significant increasing expressions of LCMR1 mRNA and protein in the 95C cells after transfection(P ＜ 0.05).Conclusions All the recombinant plasmids can be confirmed by sequencing.The 95C cells infected with lentiviral vector carrying full length LCMR1 gene express high-level LCMR1 successfully.
Keywords：Lung cancer metastasis related protein 1Lentiviral vectorVector constructionLung cancer
Publication Date：2013-01-01
Online Publishing Date：2026-09-14(First online date of this platform, not the publication date of the document)
Pages：3( 615-617 )