Video Experimental Relacionado
Updated: Jul 28, 2026
21:01
Generating Chimeric Zebrafish Embryos by Transplantation
Published on: July 17, 2009
Aislamiento del gen de la timidina quinasa del pollo por rescate de plásmidos
Nature
|May 22, 1980
Resumen
Los investigadores clonaron el gen de la timidina quinasa (tk) del pollo utilizando el plásmido bacteriano pBR322. Este gen transforma eficazmente las células tk-animales, igualando la eficiencia de los genes virales tk.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- La clonación genética.
- La expresión génica de los eucariotas.
Sus antecedentes:
- Los plásmidos bacterianos como pBR322 son herramientas esenciales para la clonación de genes.
- Los marcadores seleccionables son cruciales para identificar con éxito las células transformadas.
- El aislamiento de genes eucariotas requiere vectores de clonación eficientes.
Objetivo del estudio:
- Para utilizar el plásmido bacteriano pBR322 como vector para aislar genes eucariotas.
- Para clonar el gen de la timidina quinasa (tk) del pollo. gen.
- Para evaluar la expresión funcional del gen tk de pollo clonado en células animales.
Principales métodos:
- La clonación de genes utilizando el plásmido pBR322.
- Digestión de enzimas de restricción (EcoRI/HindIII) para la inserción de genes.
- Transformación de las células tk- animales con el plásmido recombinante.
Principales resultados:
- Se aisló con éxito el gen de la timidina quinasa (tk) de pollo como un insert de 2.2 kilobases en pBR322.
- El gen tk del pollo clonado demostró actividad funcional en la transformación de las células animales tk.
- La eficiencia de transformación fue comparable a la del gen del virus del herpes simple-1 tk.
Conclusiones:
- El plásmido bacteriano pBR322 es un vehículo viable para la clonación de genes marcadores seleccionables eucariotas funcionales.
- El gen tk de pollo clonado se puede utilizar eficazmente para la transferencia de genes y la selección en células animales.
- Este método proporciona una alternativa a los genes tk virales para la manipulación genética de las células animales.
Videos de Conceptos Relacionados
01:16In-vitro Mutagenesis
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
02:32Position-effect Variegation
In 1928, a German botanist Emil Heitz observed the moss nuclei with a DNA binding dye. He observed that while some chromatin regions decondense and spread out in the interphase nucleus, others do not. He termed them euchromatin and heterochromatin, respectively. He proposed that the heterochromatin regions reflect a functionally inactive state of the genome. It was later confirmed that heterochromatin is transcriptionally repressed, and euchromatin is transcriptionally active chromatin.
02:59Animal Mitochondrial Genetics
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
02:08Gene Conversion
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
02:30Genomic Imprinting and Inheritance
Diploid organisms inherit genetic material through chromosomes from both parents. Copies of the same gene are known as alleles. In most cases, both alleles are simultaneously expressed and allow various cellular processes to function optimally. If one of the alleles is missing or mutated, the expression of the other allele can compensate; however, this is not true for all genes.
The expression of some genes depends on which parent passed the gene to the offspring, through a phenomenon known as...
The expression of some genes depends on which parent passed the gene to the offspring, through a phenomenon known as...
02:36Chromatin Immunoprecipitation- ChIP
Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...