Cloning, Sequence Analysis And Studies Of Nucleic Acid Probe Of The TK Gene Of Infectious Laryngotracheitis Virus (ILTV) Inner Mongolia Strain

In this study,ILTV-NM98a strain and ILTV-wanggang strain were multiplied in chorioallantois.A pair of primers were devised according to the nucleic acid sequence of ILTV TK gene and the DNA of multiplied virus was used as pattern to amplify the gene of Tk by polymerase chain reaction(PCR).The product of PCR was linked with suitable plasmid.Then,the recombined plasmid was converted to Escherichia coli.The converted Escherichia coli. Was multiplied and the TK gene was cloned.The cloned TK gene was retrieved by proper restrictive hemodynamics.The retrieved TK gene was labeled by digoxin according to the kit of labeling and detection of digoxin.Then,the specificity and sensitivity of TK gene probe were detected with dot blot hybridization.The sequence of TK gene of NM98a strain was analysed.The result of the analysis of TK gene's sequence confirmed that autoploidy between TK gene of NM-98a strain and issued strain was 99.7%.The specificity of the nucleic acid probe was very strict.lt reacted positively with ILTV DNA only and it react negatively with the nuleic acid of NDV,BV and IBDV.The sensitivity of this kind of probe is very high.Ilt could even detect 20pg's ILTV DNA.