Scientia Agricultura Sinica ›› 2005, Vol. 38 ›› Issue (08): 1712-1716 .

• RESEARCH NOTES • Previous Articles    

Construction of Subtractive cDNA Libraries of Eimeria tenella to Maduramycin and Diclazuril by Suppression Subtractive Hybridization

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  1. 中国农业科学院上海家畜寄生虫病研究所
  • Received:2004-09-02 Revised:1900-01-01 Online:2005-08-10 Published:2005-08-10

Abstract: In order to clone and identify differentially expressed genes in Eimeria tenella relating to drug resistance to Diclazuril and Maduramycin, the cDNA from sporulated oocysts of the drug-sensitive strain of E.tenella was used as driver and the cDNAs from sporulated oocysts of two drug-resistant strains were used as tester to construct subtractive cDNA libraries using the technique of suppression subtractive hybridization(SSH). PCR amplification revealed that the two subtractive cDNA libraries of Maduramycin and Diclazuril-resistant strains of E.tenella contained approximated 96% and 98% recombinant clones, respectively. Plasmid inserts were PCR amplified and the lengths were 250 bp-1.0 kb. The results of dot-blot hybridization showed that six clones were over-expressed in the drug-resistant strains. Sequence analyses revealed that three and four cDNA fragments were likely to represent novel genes relating to resistance to Maduramycin and Diclazuril, respectively. These results have provided the foundation for cloning full-length genes and studying molecular mechanism relating to drug-resistance in E.tenella.

Key words: Eimeria tenella, Maduramycin, Diclazuril, Genes relating to drug resistance, Suppression subtractive hybridization, Gene cloning

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