Cargando…

A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo

We have constructed a novel tetra-promoter vector (pBVboostFG) system that enables screening of gene/cDNA libraries for functional genomic studies. The vector enables an all-in-one strategy for gene expression in mammalian, bacterial and insect cells and is also suitable for direct use in vivo. Viru...

Descripción completa

Detalles Bibliográficos
Autores principales: Laitinen, Olli H., Airenne, Kari J., Hytönen, Vesa P., Peltomaa, Erik, Mähönen, Anssi J., Wirth, Thomas, Lind, Miia M., Mäkelä, Kari A., Toivanen, Pyry I., Schenkwein, Diana, Heikura, Tommi, Nordlund, Henri R., Kulomaa, Markku S., Ylä-Herttuala, Seppo
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC549581/
https://www.ncbi.nlm.nih.gov/pubmed/15731335
http://dx.doi.org/10.1093/nar/gni042
_version_ 1782122438586269696
author Laitinen, Olli H.
Airenne, Kari J.
Hytönen, Vesa P.
Peltomaa, Erik
Mähönen, Anssi J.
Wirth, Thomas
Lind, Miia M.
Mäkelä, Kari A.
Toivanen, Pyry I.
Schenkwein, Diana
Heikura, Tommi
Nordlund, Henri R.
Kulomaa, Markku S.
Ylä-Herttuala, Seppo
author_facet Laitinen, Olli H.
Airenne, Kari J.
Hytönen, Vesa P.
Peltomaa, Erik
Mähönen, Anssi J.
Wirth, Thomas
Lind, Miia M.
Mäkelä, Kari A.
Toivanen, Pyry I.
Schenkwein, Diana
Heikura, Tommi
Nordlund, Henri R.
Kulomaa, Markku S.
Ylä-Herttuala, Seppo
author_sort Laitinen, Olli H.
collection PubMed
description We have constructed a novel tetra-promoter vector (pBVboostFG) system that enables screening of gene/cDNA libraries for functional genomic studies. The vector enables an all-in-one strategy for gene expression in mammalian, bacterial and insect cells and is also suitable for direct use in vivo. Virus preparation is based on an improved mini Tn7 transpositional system allowing easy and fast production of recombinant baculoviruses with high diversity and negligible background. Cloning of the desired DNA fragments or libraries is based on the recombination system of bacteriophage lambda. As an example of the utility of the vector, genes or cDNAs of 18 different proteins were cloned into pBVboostFG and expressed in different hosts. As a proof-of-principle of using the vector for library screening, a chromophoric Thr(65)-Tyr-Gly(67)-stretch of enhanced green fluorescent protein was destroyed and subsequently restored by novel PCR strategy and library screening. The pBVboostFG enables screening of genome-wide libraries, thus making it an efficient new platform technology for functional genomics.
format Text
id pubmed-549581
institution National Center for Biotechnology Information
language English
publishDate 2005
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-5495812005-02-26 A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo Laitinen, Olli H. Airenne, Kari J. Hytönen, Vesa P. Peltomaa, Erik Mähönen, Anssi J. Wirth, Thomas Lind, Miia M. Mäkelä, Kari A. Toivanen, Pyry I. Schenkwein, Diana Heikura, Tommi Nordlund, Henri R. Kulomaa, Markku S. Ylä-Herttuala, Seppo Nucleic Acids Res Methods Online We have constructed a novel tetra-promoter vector (pBVboostFG) system that enables screening of gene/cDNA libraries for functional genomic studies. The vector enables an all-in-one strategy for gene expression in mammalian, bacterial and insect cells and is also suitable for direct use in vivo. Virus preparation is based on an improved mini Tn7 transpositional system allowing easy and fast production of recombinant baculoviruses with high diversity and negligible background. Cloning of the desired DNA fragments or libraries is based on the recombination system of bacteriophage lambda. As an example of the utility of the vector, genes or cDNAs of 18 different proteins were cloned into pBVboostFG and expressed in different hosts. As a proof-of-principle of using the vector for library screening, a chromophoric Thr(65)-Tyr-Gly(67)-stretch of enhanced green fluorescent protein was destroyed and subsequently restored by novel PCR strategy and library screening. The pBVboostFG enables screening of genome-wide libraries, thus making it an efficient new platform technology for functional genomics. Oxford University Press 2005 2005-02-24 /pmc/articles/PMC549581/ /pubmed/15731335 http://dx.doi.org/10.1093/nar/gni042 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Methods Online
Laitinen, Olli H.
Airenne, Kari J.
Hytönen, Vesa P.
Peltomaa, Erik
Mähönen, Anssi J.
Wirth, Thomas
Lind, Miia M.
Mäkelä, Kari A.
Toivanen, Pyry I.
Schenkwein, Diana
Heikura, Tommi
Nordlund, Henri R.
Kulomaa, Markku S.
Ylä-Herttuala, Seppo
A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title_full A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title_fullStr A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title_full_unstemmed A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title_short A multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
title_sort multipurpose vector system for the screening of libraries in bacteria, insect and mammalian cells and expression in vivo
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC549581/
https://www.ncbi.nlm.nih.gov/pubmed/15731335
http://dx.doi.org/10.1093/nar/gni042
work_keys_str_mv AT laitinenollih amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT airennekarij amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT hytonenvesap amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT peltomaaerik amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT mahonenanssij amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT wirththomas amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT lindmiiam amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT makelakaria amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT toivanenpyryi amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT schenkweindiana amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT heikuratommi amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT nordlundhenrir amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT kulomaamarkkus amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT ylaherttualaseppo amultipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT laitinenollih multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT airennekarij multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT hytonenvesap multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT peltomaaerik multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT mahonenanssij multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT wirththomas multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT lindmiiam multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT makelakaria multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT toivanenpyryi multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT schenkweindiana multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT heikuratommi multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT nordlundhenrir multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT kulomaamarkkus multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo
AT ylaherttualaseppo multipurposevectorsystemforthescreeningoflibrariesinbacteriainsectandmammaliancellsandexpressioninvivo