Cloning and characterization of Rab and Ran/TC4 cDNA clones in Trichinella spp. |
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| Authors: | Esperanza Rodríguez Pedro Bonay Manuel Fresno Teresa Gárate |
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| Institution: | (1) Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III, Centro Nacional de Microbiología, Majadahonda, E-28220 Madrid, Spain e-mail: tgarate@isciii.es, Tel.: +3415097901 (x3603), Fax: +3415097966, ES;(2) Centro de Biología Molecular, Trescantos, E-28046 Madrid, Spain, ES |
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| Abstract: | The cloning and characterization of seven Rab and three Ran/TC4 partial cDNA sequences in both cystic (Trichinella spiralis and T. britovi) and noncystic species (T. pseudospiralis) are reported. These molecules were cloned by rapid amplification of cDNA ends via polymerase chain reaction (RACE-PCR),
using cDNA from the aforementioned Trichinella spp. coupled to the AP1 adaptor. As primers, AP1 and 5B (derived from the WDTAGQE sequence of region 2 specific for Rab and
Ran proteins) sequences were included in the PCR. The cloned cDNAs were sequenced and characterized by both Southern-blot
and Northern-blot analysis. Trichinella spp. Rab- and Ran-like molecules showed divergences in both the nucleotide and the deduced amino acid sequences as compared
with the corresponding homologues previously described in other organisms. In addition, differences were observed among the
Trichinella species, mainly between the cystic and the noncystic species, in both DNA restriction-enzyme polymorphism and expression
of the six GTPases isolated.
Received: 12 November 1998 / Accepted: 3 February 1999 |
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