Asunto(s)
Ciclo Celular , Expresión Génica/fisiología , Proteínas Represoras/biosíntesis , Tetraciclina/farmacología , Animales , Línea Celular , Clonación Molecular/métodos , Citomegalovirus/genética , Expresión Génica/efectos de los fármacos , Genes Bacterianos , Genes Reporteros , Vectores Genéticos , Globinas/biosíntesis , Humanos , Intrones , Luciferasas/biosíntesis , Mamíferos , Plásmidos , Regiones Promotoras Genéticas , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Represoras/genética , Simplexvirus/genética , Resistencia a la Tetraciclina/genética , Transactivadores/metabolismo , Transfección/métodosRESUMEN
Of the wide variety of regulatory systems that have been developed to control gene expression in higher eukaryotes, those utilizing elements from prokaryotic systems presently achieve the highest specificity. In particular, systems based on the repressor/operator elements of an Escherichia coli tetracycline resistance operon appear broadly applicable. During the past year, the usefulness of these systems has been demonstrated not only at the cellular level, but also at the organismal level (i.e. in transgenic animals and plants).
Asunto(s)
Expresión Génica , Animales , Animales Modificados Genéticamente , Escherichia coli/genética , Células Eucariotas , Regulación de la Expresión Génica , Humanos , Operón , Plantas Modificadas Genéticamente , Resistencia a la Tetraciclina/genética , Transcripción GenéticaRESUMEN
The coding sequence of the Photinus pyralis luciferase (Luc), as contained in the previously described expression vector, pUHC13-1, was modified such that several commonly used restriction sites in the luc gene were removed without changing the primary sequence of the protein. In addition, a Kozak sequence upstream from the ATG start codon was introduced. The resulting plasmid, pUHC131-1, produced increased amounts of Luc activity in HeLa and NIH3T3 cells. The modified luc gene was inserted in both orientations into the multiple cloning site of a commonly used cloning vector. This facilitates transfer of luc into other expression units.
Asunto(s)
Escarabajos/enzimología , Genes de Insecto , Genes Reporteros/genética , Vectores Genéticos , Luciferasas/genética , Animales , Secuencia de Bases , Línea Celular , Clonación Molecular , Escarabajos/genética , Células HeLa , Humanos , Luciferasas/biosíntesis , Datos de Secuencia MolecularRESUMEN
Regulated gene expression systems for the study of gene function in prokaryotes and yeast have been available for several years. However, comparable systems in higher organisms are more complex and problematic. Recently, regulatory proteins from distant species have been used to establish highly specific control systems in eukaryotic cells. This is possible because their action can be modulated by effectors that are inert to the physiology of the organism or cell and therefore do not elicit pleiotropic effects. Such monospecific regulatory circuits open up new possibilities for the study of gene function in vivo.
