Gen de control interno VpEf1α en Vasconcellea pubescens (chamburo)

Tiffany Yiselle Cevallos Vilatuña; Karen Alejandra Garzón Salazar; Fabio Marcelo Idrovo Espín

doi:10.59410/RACYT-v08n01ep01-0103

Autores/as

Tiffany Cevallos Vilatuña Universidad de las Américas
Karen Alejandra Garzón Salazar Universidad de las Américas
Fabio Marcelo Idrovo Espín Central University of Ecuador

DOI:

https://doi.org/10.59410/RACYT-v08n01ep01-0103

Palabras clave:

Housekeeping gene, V. pubescens, expresión, secuenciación, filogenia

Resumen

Los genes conocidos como “housekeeping” controlan o regulan procesos celulares básicos y permanecen activados siempre, independientemente de las condiciones experimentales o entre las células de diferentes tejidos. Vasconcellea pubescens, es una especie ampliamente distribuida en América del Sur y pertenece a la familia Caricaceae al igual que la papaya. En primer lugar, se diseñaron primers para el gen EF1α en base al genoma de Carica papaya y Arabidopsis thaliana. Después, plántulas de V. pubescens se sometieron a tres temperaturas diferentes. La cuantificación de la expresión relativa del gen se realizó por densitometría. Finalmente, los fragmentos obtenidos de la RT-PCR se secuenciaron por Secuenciación Sanger de segunda generación y los análisis bioinformáticos se realizaron con MEGA X mientras que los análisis estadísticos se realizaron con RCommander. Se obtuvo un fragmento de 173 pb que se denominó VpEF1α. La secuencia de nucleótidos y la traducción a aminoácidos resultaron ser muy similares al compararlas con secuencias Ef1α conocidas de otras especies vegetales. A partir de la filogenia realizada con la proteína predicha, VpEF1α se agrupó en un solo clado con secuencias de álamo, cacao y papaya, todas ellas arbóreas, mientras que Arabidopsis y tabaco se ubicaron en otro clado. La expresión del gen VpEF1α fue similar en las tres temperaturas evaluadas cumpliendo el requisito de que no cambie su expresión a diferentes condiciones experimentales. Se describió de esta forma un gen tipo EF1α en V. pubescens (chamburo) que podría ser utilizado como gen control interno o housekeeping en estudios futuros.

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