Expression of t-PA(1354-1802) and Preparation of Its antiserum

[Objective]To prepare t-PA(1354-1802) antiserum and measure its titer,so as to lay a foundation for the deep studies on gene function of t-PA.[Method]The 449 bp fragment of t-PA was obtained by gene cloning techniques,and then recombined into pET-32a(+) vector.The recombinant vector was confirmed by PCR and DNA sequence analysis.The positive recombinant plasmid was transformed into BL21(DE3) and then induced with IPTG for expression.[Result]An expected specific band of about 37 kD was determined by SDS-PAGE.The fusion protein was purified and then used to immunize the New Zealand rabbits,and protein t-PA(1354-1802) antiserum was prepared.Western blot analysis showed that the antiseum could bind to standard of t-PA protein.The ELISA titer of the rabbit i-t-PA(1354-1802) antiserum was 1∶ 128 00 approximately.[Conclusion]t-PA(1354-1802)antiserum is successfully prepared,which lays foundation for the in-depth study of t-PA gene.