ISSN 2736-1756
Advanced Journal of Microbiology Research ISSN 2241-9837 Vol. 13 (3), pp. 001-006, March, 2019. © International Scholars Journals
Full Length Research Paper
Study on differential display gene expression of Eimeria tenella multiple resistance strain isolated from Tangshan in chicken
Zhang Yanying, Zhang xiangzhai, Jia Qinghui, Li Yunyu, Li Peiguo*, Zhang Wenxiang and Chen Juan
Key Laboratory of Preventive Veterinary Medicine in Hebei Province,
Hebei Normal University of Science and Technology, Qinhuangdao, 066004 China.
Accepted 15 January, 2019
Abstract
Total RNA of Eimeria tenella drug-resistant strain from Tangshan was extracted with Trizol. Differential-display reverse transcription- polymerase chain reaction (DDRT-PCR) was established by 3 anchored primers and 20 arbitrary primers. The products of polymerase chain reaction (PCR) were analyzed on the denaturing polyacrylamide gels by silver-straining. Ten differential bands were excised from the gels and reampilfied with the same sets of primers. The products were purified and ligated with PMDTM18-T Vector, and then the dot-blot hybridization, sequence analysis and homology comparison. The results showed that through comparison of the nucleotide acid sequence, the similarity was 99% among the sequence S116 from mRNA of Tangshan multiple-resistant strain with the sequence 882 bp lengths in the first chromosome of E.tenella in Genebank and Sanger, which was an unknown protein. This study paved the way for cloning the full-length cDNAs (Complementary Deoxyribonucleic acid) and finding the molecular mechanism about the drug-resistance of E.tenella.
Key words: Chicken, Eimeria tenella, mRNA differential display polymerase chain reaction (PCR), denaturing polyacrylamide gel, gene cloning.