Yunus, Muchammad (2012) Isolation of Total RNA of Sporozoite Membrane Protein of E.Tenella as Principle of Fragment Gene Amplification PCR Technique. Proceedings International Seminar “A Role of The Veterinarian on The Global Health Challenge. pp. 27-30. ISSN 978-602-18660-8-5

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Abstract

The use of sporozoite can be used to increase chicken resistance on disease and/or infection on commercial farm. Initially, protein of sporozoite membrane of E. tenella was isolated and characterized expression of code gene protein of sporozoite membrane as principle of recombinant technique. The first step was to performe isolation and characterisation of protein of sporozoite membrane by extraction technique of protein of sporozoitmembraneof E. tenella using SDS-PAGEand Dot Blot.Theresultof isolation of protein of sporozoite membrane was obtained optical density (OD) with indirect ELISA. Isolation RNA total of protein of sporozoitemembraneshowed band of rRNA about 23S and 1 6S of marker band, which bands of mRNA was seen smear among bands of rRNA. Molecular characterisation was performed using RT-PCR and genetic analyses were done to PCR product. The course of isolation of cDNA from RNA total and mRNA also the result of nucleotide sequence analyses of protein of sporozoite membrane showed fragmenlenght of 480 bp. Gene fragmen amplification of protein of sporozoite membrane E. lene/la of PCR product and based on the result of DNA electrophoresis using specific primer forward and primer reverse produced PCR product in fragmenlenght at 366 bp. sporozoite membrane protein, E. tenella

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