Published June 2011 | Version v1
Miscellaneous

Subcloning of phospholipase C of Clostridium perfringens as fusion protein with green fluorescent protein

Description

The gene that codes for Clostridium perfringens phospholipase C was amplified to clone it into the plasmid vector pGEX, which has a sequence that codes for the green fluorescent protein. The recombinant plasmid was used to transform competent cells of Escherichia coli BL-21. The clones obtained were selected by the resistance to ampicillin conferred by the β -lactamase present in the plasmid. Later the clones were characterized to confirm in them the presence of the gene that codes for the phospholipase C of Clostridium perfringens. The production of the fusion protein Cp-PLC-GFP (phospholipase C from Clostridium perfringens-green fluorescent protein) was allowed by the clones, which, being marked, will be a useful tool in the determination of the intracellular transit pathway of this toxin and the target tissues of it. (author)

Availability note (English)

Available from Biblioteca Luis Demetrio Tinoco, Universidad de Costa Rica

Abstract (Spanish)

El gen que codifica para la fosfolipasa C de Clostridium perfringens fue amplificado para clonarlo en el plasmido vector pGEX, el cual posee una secuencia que codifica para la proteina verde fluorescente. El plasmido recombinante fue utilizado para transformar celulas competentes de Escherichia coli BL-21. Los clones obtenidos fueron seleccionados mediante la resistencia a la ampicilina conferida por la β-lactamasa presente en el plasmido. Posteriormente los clones fueron caracterizados para confirmar en ellos la presencia del gen que codifica para la fosfolipasa C de Clostridium perfringens. La produccion de la proteina de fusion Cp-PLC-GFP (fosfolipasa C de Clostridium perfringens-proteina verde fluorescente) fue permitida por los clones, la cual al estar marcada, sera una herramienta util en la determinacion de la via de transito intracelular de esta toxina y los tejidos diana de la misma. (autor)

Additional details

Additional titles

Original title (Spanish)
Subclonaje de la fosfolipasa C de Clostridium perfringens como proteina de fusion con la proteina verde fluorescente

Publishing Information

Imprint Place
San Jose (Costa Rica)
Imprint Pagination
42 p.

INIS

Country of Publication
Costa Rica
Country of Input or Organization
Costa Rica
INIS RN
49086744
Subject category
S60: APPLIED LIFE SCIENCES;
Resource subtype / Literary indicator
Numerical Data, Thesis, Non-conventional Literature
Descriptors DEI
BACTERIAL DISEASES; BIOTECHNOLOGY; CHROMATOGRAPHY; CLONING; CLOSTRIDIUM PERFRINGENS; ELECTROPHORESIS; EXPERIMENTAL DATA; GENETICS; MOLECULAR BIOLOGY; MOLECULES; POLYMERASE CHAIN REACTION
Descriptors DEC
BACTERIA; BIOLOGY; CLOSTRIDIUM; DATA; DISEASES; GENE AMPLIFICATION; INFECTIOUS DISEASES; INFORMATION; MICROORGANISMS; NUMERICAL DATA; SEPARATION PROCESSES

Optional Information

Notes
Tabs., figs., refs.