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Biotechnol Lett ; 35(1): 21-7, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22955677

RESUMEN

The expression enhancement by cytomegalovirus promoter and different intron A (IA) variants were evaluated in CHO-K1, HepG2, HEK-293 and COS-7 cells by assessing the levels of luciferase activity. This data along with mRNA levels measurement indicated that the construct harboring an IA variant with a 200-nucleotide deletion (Δ200) had the greatest impact on increasing luciferase expression among all constructs evaluated. Based on these results, we redesigned pCMV-IA variants and cloned them into plasmids expressing a humanized antibody. These plasmids were then used to transfect CHO-K1 cells. Production of the antibody was not augmented with the Δ200 promoter variant. The 600-nucleotide deletion (Δ600) and whole IA promoter variants expressed similar levels of the recombinant protein. These data indicate that the IA-based enhanced expression of transgenes depends on a small region within the intron.


Asunto(s)
Citomegalovirus/genética , Intrones , Proteínas Recombinantes/biosíntesis , Transgenes , Animales , Biotecnología , Células CHO , Células COS , Chlorocebus aethiops , Cricetinae , Cricetulus , Expresión Génica , Células HEK293 , Humanos , Luciferasas/análisis , Luciferasas/genética , Luciferasas/metabolismo , Regiones Promotoras Genéticas , ARN Mensajero/análisis , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/análisis , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
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