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Designing, cloning and amplification of pDream2.1/MCS/CII-6 recombinant plasmid which includes a mexican scorpion Centruroides limpidus limpidus CII-6 gene
* 1 , 2 , 3 , 4 , 1 , * 1
1  Laboratorio de Biología Molecular, Centro de Investigación y Estudios Avanzados en Salud Animal (CIESA), Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma del Estado de México (UAEM), Carretera Toluca - Atlacomulco Km. 14.5, San Cayetano
2  Laboratorio de Biología Molecular y Bioquímica, Centro Universitario de los Lagos, Universidad de Guadalajara, Avenida Enrique Díaz de León 1144, Paseos de La Montaña, Lagos de Moreno, Jalisco C.P. 47460.
3  Hospital Veterinario Pequeñas Especies, Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma del Estado de México (UAEM), Jesús Carranza No. 203, Col. Universidad, Toluca, Estado de México C.P. 50130.
4  Laboratorio de Biotecnología, División de Ingeniería en Biotecnología, Universidad Politécnica del Valle de Toluca, Carretera Almoloya de Juárez, Km 5.6, Santiaguito Tlalcilalcali, Almoloya de Juárez, Estado de México, C.P. 47460.

Abstract:

Biologically active molecules present in the venom of several species of scorpion, have shown potential against various diseases, including cancer. It has been reported that several toxins can block ion channels present in the membrane of several cancer cell lines, through action potential, altering their cell function, cell cycle arresting, and inducing apoptosis death pathway. The use of the sequence of Cll-6 gene present in the genome of the Mexican scorpion Centruroides limpidus limpidus encoding a beta-toxin-locker of Na + channels by action potentials is proposed. This molecule has not been evaluated to determine if it has potential as an anti-cancer agent. In this paper the in silico design of recombinant molecule pDream2.1/MCS/CII-6, including the CII-6 gene into the polylinker site and identification by molecular biology techniques. Our results confirm a fragment of 316 base pairs, by digestion with BamH1 and Kpn1 enzymes, PCR, and the sequenced of amplicon, the alignment with CII-6 sequence reported in the GenBank database, showed 99.6% similarity and identity. The recombinant plasmid, could be used to assess potential as anti-having cancer agent on several cancer cell lines.

Keywords: CII-6 toxin; Centruroides limpidus limpidus; pDream2.1/MCS; Cloning; Transformation; Escherichia coli; canal ionic; cancer
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