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Study on amplification and its optimum reaction conditions of E2 gene of Hog Cholera Virus Guangxi prevalence strain

Author: WeiDongPing
Tutor: LuQinZhang；LuoTingRong
School: Guangxi University
Course: Preventive Veterinary Medicine
Keywords: Reverse transcription - polymerase chain reaction Classical swine fever virus epidemic strains in Guangxi E2 gene The optimum reaction conditions
CLC: S852.651
Type: Master's thesis
Year: 2001
Downloads: 72
Quote: 0
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Abstract


This paper presents the HCV-specific primers A1/A2 directly from parts of Guangxi 58 suspected swine fever epidemic materials detection, of which 34 were positive, the positive rate was 58.32% . Of classical swine fever virus nucleic acid sequence based on the published design and synthesis of the four pairs of EP1/EP4 EP2/EP3 EP3/EP7 and EP5/EP6 to the cited the the matter EP1/EP4 and EP2/EP3 or EP3/EP7 and EP5/EP6 paired by RT-PCR technology was divided into two phases of classical swine fever virus E2 gene was amplified , and the reaction conditions were optimized . : The primer EP1/EP4 and EP2/EP3 only amplified HCLV ( HCV ) and Shimen virulent and can not be amplified the epidemic strains Guangxi : EP3/EP7 EP5/EP6 can successfully amplified Guangxi epidemic strain and two other strains . The reaction conditions required for reverse transcription of total RNA 10μ1 ( approximately 20 microg ) ; PCR amplification , cDNA used in an amount of 12.5μl, positive , negative primer with 50 pmol each the EP3/EP7 and EPS/EP6 of primers , respectively, PCR amplification for 35 cycles and 61 ° C annealing temperature to 57 ° C t , PCR product amount and the amplification specificity sentences optimum level . E2 gene of classical swine fever virus epidemic strains in Guangxi successfully amplified and the establishment of optimal reaction conditions lay the foundation for further the E2 gene cloning and sequence analysis of its in-depth study .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Livestock Virology > Classical swine fever virus
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