摘要
利用RACE技术从19日龄的日本血吸虫童虫中扩增了1个Wnt家族基因,并对其进行了生物信息学分析。同源性分析表明,该基因为日本血吸虫新基因,完整开放阅读框(ORF)长1896 bp,编码631个氨基酸,理论分子质量为73.3 ku。该基因编码的氨基酸序列具有Wnt家族蛋白的典型特征,与人、鼠Wnt10a的氨基酸序列同源性均为26%,推测为血吸虫的Wnt10a基因,命名为Sjwnt10a(Gen BanK登录号DQ643829)。利用实时荧光定量PCR分析该基因在日本血吸虫不同发育阶段虫体中的表达情况,结果显示该基因在19日龄的日本血吸虫童虫中表达量最高,在14日龄童虫和44日龄雄虫中也有表达,但分别仅为19日龄童虫表达量的8.8%和5.0%,而在31日龄成虫和44日龄雌虫中没有检测到该基因。结果提示,童虫差异表达的Sjwnt10a基因可能对童虫的生长、发育至关重要。
Based on an EST from 19-day-old schistosomula identified in our laboratory, a novel gene was cloned with RACE technique and analysed with bioinformatics. The cloned gene had a complete ORF containing 1 896 nucleotides, and encoding 631 amino acid with 73.3 ku of molecular mass. Sequence analysis showed that the gene had typical characteristics encoding Wnt protein family and shared 26 % similarity to mouse and human Wnt10a, hence, the gene was named as SjwntlOa（GenBank Accession Number： DQ643829）. Real-time fluorescent quantitative PCR revealed that the mRNA level of SjwntlOa was the highest in 19-day-old schistosomula, and lower in 44-day-old male worms and 14-day-old schistosomula, which were only 8.8% and 5.0% of that of the 19-day-old schistosomula,respectively, and there were no expression of the gene in 31-day-old adult worms and 44-day-old female worms. The results suggested that Sjwnt10a is a stage-and-gender differentially expressed gene which may play an important role in the development of schistosomula.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第2期93-97,共5页
Chinese Veterinary Science
基金
国家自然科学基金资助项目(30600450
30471299和30671581)
