摘要
以从大白菜温敏雄性育性转化相关基因差异表达的分析中获得的EST-58为标签,采用电子克隆的方法对其进行延伸,并对电子克隆的结果进行RT-PCR验证,结果表明:电子克隆到1个由1197bp核苷酸组成的序列,该序列具有完整的开放阅读框架(ORF),编码蛋白为292个氨基酸。经RT-PCR扩增,连续样品间EST-58表达的带型变化趋势与cDNA-AFLP的差异显示情况一致,且RT-PCR获得的序列与电子克隆的序列一致性达98%。用该序列在GenBank数据库中进行blastX同源性分析,确定该序列为大白菜的XET基因(GenBank登陆号为EU579461)。
The EST-58 related to the fertility changeover in the thermo-sensitive CMS （TsCMS7311） of Chinese cabbage （Brassica campestris L.ssp.pekinensis） was amplified by in silico cloning.And the cDNA sequence had been confirmed by RT-PCR.The result indicated that a new sequence with 1197 bp length was cloned,which contained an integrated ORF,encoding 292 amino acids.The trend of EST-58 expression by RT-PCR was very identity to the expression by cDNA-AFLP,and their sequences have 98% consistency.As showed that the result of in silico cloning was right.The amino acid sequence showed highly comparability with xyloglucan endotransglycosylase （XET） from GenBank.It indicated that the new sequence was XET gene （GenBank accession,EU579461） of Chinese cabbage.
出处
《中国蔬菜》
北大核心
2010年第24期19-24,共6页
China Vegetables
基金
国家自然科学基金(30871717)
国家科技支撑计划(2009BADB8B03)
