摘要
从伴刀豆球蛋白 (ConA)刺激的鸡脾细胞中扩增出鸡白细胞介素 1β(ChIL 1β)编码区基因 ,通过与GenBank上登录的ChIL 1β序列 (Y15 0 0 6 )进行比较。结果显示 ,在ChIL 1β基因编码区的 10 4位、30 6位、387位和 4 32位的核苷酸发生了变化 ;相应的氨基酸在 35位也发生了变化。虽然 30 6位、387位和 4 32位的核苷酸发生了变化 ,但没有引起氨基酸变化 ,说明这一区域的氨基酸具有一定的保守性。从氨基酸的性质上看 ,35位 (Asp→Ala)由酸性氨基酸变为中性氨基酸 ,这可能是同一种属的不同品种间的正常变化。以鸡痘病毒 (FPV )为活载体 ,构建了表达ChIL 1β的重组鸡痘病毒rFPV IL 1β。应用XTT/PMS方法检测rFPV IL 1β感染的成纤维细胞 72h后表达ChIL 1β的生物活性 ,效价为 1.0× 10 5U /mL ,证明FPV能有效地表达ChIL 1β。
The cDNA of the chicken interleukin-1β (ChIL-1β) was cloned from ConA-stimulated chicken spleen cells by RT-PCR. Compared with the published ChIL-1β sequence in the GenBank(accession No.Y15006), nucleotide changes in the cloned ChIL-1β sequence were found at nt 104(A→G),306(C→T),387(G→T)and 432(C→A). And the corresponding change of amino acid was found at amino acid 35 (Asp→Ala) of putative polypeptide. The nucleotide variation at nt 306(C→T),387(G→T)and 432(C→A) did not result in amino acid change. Then a recombinant fowlpox virus(rFPV) expressing ChIL-1β was constructed and the biologic activity of the recombinant ChIL-1β expressed by rFPV in chicken embryo fibroblasts in vitro using XTT/PMS was determined. The result demonstrated that FPV can express effectively ChIL-1β.
出处
《中国兽医科技》
CSCD
北大核心
2004年第1期12-16,共5页
Chinese Journal of Veterinary Science and Technology
基金
自然科学基金项目 (BK2 0 0 14 15 )
