University of Tehran Iranian Journal of Veterinary Medicine 2251-8894 7 1 2013 04 01 The mRNAexpression study on small amount of Theileria annulata lymph node biopsy sample using SMART-cDNA technology 15 22 32019 10.22059/ijvm.2013.32019 EN Nastaran Sadr Shirazi Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran Parviz Shayan Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran; Institute of Molecular Biological System Transfer (MBST), Tehran, Iran Berigiteh Eckert Institute of Molecular Biological System Transfer (MBST), Tehran, Iran Elaheh Ebrahimzadeh Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran Sedigheh Jafari Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran Journal Article 2012 05 15 BACKGROUND: A major issue in many gene expression<br />studies utilizing small amount of biological materials is the<br />limited quantity of RNApurified from clinical samples, which is<br />often used for RT-PCR or standard Northern blot analysis.<br />OBJECTIVES: The SMART cDNA synthesis method and subsequent<br />SMART-cDNA-PCR technique was used to analyse 3<br />genes in macroschizonts of Theileria annulata in small lymph<br />node biopsy material. METHODS: The SMART-cDNA of TaSp<br />gene was cloned in pTZ57R/T-vector and sequenced. We focused<br />on genes encoding surface proteins TaSp, TaD and HSP70.<br />RESULTS: Our results showed that SMART cDNA dependably<br />reproduces the expression profile found in messenger RNA. The<br />RT-SMART-PCR showed the amplification of the processed<br />mRNAs. The sequencing analysis showed that the amplified<br />cDNA was coded for TaSp protein in Theileria annulata.<br />CONCLUSIONS: It was concluded that the SMART PCR<br />technique is practical for amplification of complete sequence of<br />mRNAs in the form of cDNAs, and therefore for gene expression<br />studies if only small amounts of starting material are available. https://ijvm.ut.ac.ir/article_32019_3e488d7a07aad80a635656ada23e1571.pdf