The effect of various introns and transcription terminators on the efficiency of expression vectors in various cultured cell lines and in the mammary gland of transgenic mice

Autor: Philippe Bolifraud, Marie-Claire Théron, Monique Béarzotti, Louis-Marie Houdebine, D Petitclerc, Joe Attal, Guy Kann, H. Pointu, Marie-Georges Stinnakre, Claudine Puissant
Přispěvatelé: ProdInra, Migration, Unité de biologie cellulaire et moléculaire, Institut National de la Recherche Agronomique (INRA), Station de virologie et d'immunologie, Unité de recherche Génétique Biochimique et Cytogénétique (LGBC)
Rok vydání: 1995
Předmět:
Transcription
Genetic
[SDV]Life Sciences [q-bio]
viruses
Cytomegalovirus
Gene Expression
Simian virus 40
Applied Microbiology and Biotechnology
Mice
0302 clinical medicine
Gene expression
Promoter Regions
Genetic
ComputingMilieux_MISCELLANEOUS
Terminator Regions
Genetic
0303 health sciences
Expression vector
biology
General Medicine
Milk Proteins
[SDV] Life Sciences [q-bio]
Enhancer Elements
Genetic
030220 oncology & carcinogenesis
Female
Whey Acidic Protein
Rabbits
Biotechnology
DNA
Complementary
Genetic Vectors
Bioengineering
Mice
Transgenic
Transfection
Cell Line
03 medical and health sciences
Capsid
Mammary Glands
Animal
Complementary DNA
Animals
Humans
RNA
Messenger
Enhancer
Gene
030304 developmental biology
VECTEUR
Intron
Promoter
Molecular biology
Introns
Growth Hormone
biology.protein
Capsid Proteins
Cattle
Zdroj: Journal of Biotechnology
Journal of Biotechnology, Elsevier, 1995, 40, pp.169-178
ISSN: 0168-1656
Popis: Various combinations of promoters, introns and transcription terminators were used to drive the expression of bovine growth hormone (bGH) cDNA in different cell types. In constructs containing the human cytomegalovirus (hCMV) promoter and the SV40 late genes terminator, the intron from SV40 genes (VP1) was much more efficient, than the intron from the early genes (t). The synthetic intron SIS generated by the association of an adenovirus splice donor and an immunoglobulin G splice acceptor showed the highest activity. The respective potency of these introns was similar in several mammalian (CHO, HC11 and COS) and fish (TO2 and EPC) cells. The rabbit whey acidic protein (WAP) gene promoter was highly efficient to drive the expression of bGH gene in the HC11 mammary cell lines. In contrast, the bGH cDNA under the control of the same promoter was much less efficiently expressed when the SV40 VP1 intron and transcription terminator were used. The rabbit WAP gene and the human GH gene terminators did not or only moderately enhanced the expression of the construct WAP bGH cDNA. Introduction of a promoter sequence from the mouse mammary tumor virus (MMTV) LTR in the VP1 intron increased very significantly the expression of the WAP bGH cDNA. Although several of these vectors showed high potency when expressed stably in HC11 cells, all of them were only moderately efficient in transgenic mice. These data indicate that the VP1 and the SIS introns may be used to express foreign cDNAs with good efficiency in different cell types. The addition of an enhancer within an intron may still reinforce its efficiency. However, transfection experiments, even when stable expression is carried out, are poorly predictive of the potential efficiency of a vector in transgenic animals.
Databáze: OpenAIRE
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