The mRNAexpression study on small amount of Theileria annulata lymph node biopsy sample using SMART-cDNA technology

Authors

1 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran

2 Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran,Tehran, Iran; Institute of Molecular Biological System Transfer (MBST), Tehran, Iran

3 Institute of Molecular Biological System Transfer (MBST), Tehran, Iran

Abstract

BACKGROUND: A major issue in many gene expression
studies utilizing small amount of biological materials is the
limited quantity of RNApurified from clinical samples, which is
often used for RT-PCR or standard Northern blot analysis.
OBJECTIVES: The SMART cDNA synthesis method and subsequent
SMART-cDNA-PCR technique was used to analyse 3
genes in macroschizonts of Theileria annulata in small lymph
node biopsy material. METHODS: The SMART-cDNA of TaSp
gene was cloned in pTZ57R/T-vector and sequenced. We focused
on genes encoding surface proteins TaSp, TaD and HSP70.
RESULTS: Our results showed that SMART cDNA dependably
reproduces the expression profile found in messenger RNA. The
RT-SMART-PCR showed the amplification of the processed
mRNAs. The sequencing analysis showed that the amplified
cDNA was coded for TaSp protein in Theileria annulata.
CONCLUSIONS: It was concluded that the SMART PCR
technique is practical for amplification of complete sequence of
mRNAs in the form of cDNAs, and therefore for gene expression
studies if only small amounts of starting material are available.

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