Screening of Multiple
Drug Resistant Genes of
Eimeria tenella Infesting
Chicken in China
Yanying Zhang, Qinghui Jia,
Xiangzhai Zhang, Yunyu Li, Peiguo Li*, Wenxiang Zhang, Juan Chen and
Wenjin Zhu
Key Laboratory of Hebei
Province, Preventive Veterinary Medicine, Hebei Normal University of
Science & Technology, Qinhuangdao, China, 066004 *Corresponding
author:
lipg6205@yahoo.com.cn
Abstract
Total RNA of
Eimeria tenella drug-resistant strains
from Qinhuangdao, Tanshan and Shijiazhuang were extracted with Trizol.
DDRT-PCR was established by 3 anchored primers and 20 arbitrary primers. The
products of PCR were analyzed on the denaturing polyacrylamide gels by
silver-straining. Twenty-six differential bands were excised from the gels and
reampilfied with the same sets of primers. The products were purified, and 12
differential bands (20~50ng) were recuperated. Twelve plasmid cloning
identification differential bands were identified by the dot-blot hybridization.
The results showed that 3 bands were positive. And then the positive cloning was
sequenced and compared to homology. The results showed that through comparison
of the nucleotide acid sequence, the similarity was 99% among the sequence S116
from mRNA of Shijiazhuang
multiple-resistant strain with the sequence 882bp length in the first chromosome
of E. tenella in Genbank and Sanger,
which was an unknown protein. The sequence of T311 and Q19 from mRNA of
Tangshan and Qinhuangdao multiple-resistant strains were
unknown sequences. These novel cDNA fragments associated with
drugresistance might be
involved in the process of the drug resistances of
E. tenella.