Phylogenic Analysis and Expression the Thymidine Kinase Gene of Infectious Laryngotracheitis Virus

• Main Authors: Hui-Chen Huang, 黃慧禎
• Other Authors: Maw-Yeong Lin
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/32831927271949685749

碩士 === 國立屏東科技大學 === 獸醫學系 === 94 === Infectious laryngotracheitis ( ILT ) is caused by infectious laryngotracheitis Herpesvirus ( ILTV ), which induces upper respiratory tract syndromes and decreases egg production with high mortality. Generally ILT is well controlled by using of live attenuated vaccine. ILTV has thymidine kinase ( TK ) gene which has been shown is associated with its virulence and reactivation from the latent state. This study is to search for variation of ILTV-TK gene, and using a pair of high sensitivity and specificity primer is chosed to amplify the full length ( 1092 bp; nt1-nt 1092 ) TK gene of 11 variant vaccine strains and 4 field isolates by polymerase chain reaction and then cloning and sequencing, with download nucleotide of 4 ILTV-TK in GenBank in comparison. it in similarities nucleotide comparison among 19 ILTV isolates ranged from 99.1 ％-100 ％, amino acid comparison ranged from 97.8 ％-99.7 ％, the nucleotide variation among 19 ILTV isolates is reasonably low. The TK fusion protein of the field isolate will be expressed in E.coli system, the recombinant protein with molecular weight of 60 kDa was obtained and western blot analysis demonstrated the specificity of expressed protein.