Temporal quantification of Ts43 gene expression of Trichinella spiralis using real-time RT-qPCR
2011
Abstract Determination of the transcription level of Trichinella spiralis ( T. spiralis ) Ts43 gene is essential for understanding its poorly explained role in different developmental stage. Herein we report on measurement of T. spiralis Ts43 mRNA by adapting a relative quantitative real-time reverse transcription and polymerase chain reaction (RT-PCR). Total RNA was isolated from the samples of adult worms, newborn larvae and muscle larvae of T. spiralis and Ts43 mRNA was quantified by real-time RT-PCR using an internally calibrated standard curve constructed with the recombinant Ts43 and 18S rRNA plasmid. The results indicated that the Ts43 gene was expressed at any stage and the expression levels in adults and newborn larvae were lower than those in muscle larvae in general. The expression of the Ts43 gene increased from 18 days post-infection (p.i.), reached a peak at 22–30 days p.i., and then decreased to a low level by 58 days p.i. but it was still higher than that in adults and newborn larvae. The study provided the first quantitative of Ts43 mRNA expression in T. spiralis for further studies of Ts43 function.
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