[Tat transduction peptide displayed on the nucleocapsid improved the baculovirus transduction of mammalian cells].
In order to improve the transduction efficiency of insect baculovirus in mammalian cells, we constructed two recombinant baculoviruses, AcRed-tat and AcRed. Both viruses expressed red fluorescence protein gene (dsRed) as a reporter in mammalian cell lines. AcRed-tat also contained the coding sequence of HIV-1 Tat transduction peptide fused with viral major capsid protein gene vp39 and enhanced green fluorescence gene (egfp) driven by virus polyhedrin promoter. It expressed the Tat fusion protein in infected insect cells, which was incorporated into the nucleocapsids of progeny virus. As a control, AcRed had the fusion gene of vp39 and egfp driven by polyhedrin promoter. Flow cytometry analysis demonstrated that although similar level of red fluorescence was produced in HEK23 cells transduced by the two recombinant viruses, significantly higher red fluorescence level was seen in CHO and Vero cells transduced by AcRed-tat than that by AcRed. These results suggested that Tat transduction peptide might improve the baculovirus-mediated gene expression in some mammalian cells. Our work provided a new approach to improve baculovirus as a gene delivery vector for mammalian cells.
Li
Li
L
Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
Wang
Xin
X
Yin
Juan
J
Zhong
Jiang
J
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D015696:Gene Products, tat / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D019251:Nucleocapsid / Q000235:genetics; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
D015696:Gene Products, tat; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C088682:VP39 protein, Vaccinia virus; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins