家蚕丝腺组织Actin3引物非特异性扩增片段的鉴定及A3引物扩增条件的优化

Identification of the A3 Non-specific Amplification Bands from the Silk Gland of Bombyx mori and the Optimization of the A3 Primer Amplyfication Conditions

肌动蛋白基因3(Actin3)是家蚕分子生物学研究中检测组织cDNA模板质量常用的标志基因之一。我们在用实验室通用的Actin3引物扩增检测家蚕不同茧色品种(白血白茧品种夏芳、金色茧品种金一、黄血白茧品种03-000、锈色茧品种03-520)中肠、血液、丝腺组织cDNA时,发现以不同茧色品种丝腺组织cDNA为模板都能扩增出一条非特异性的小片段,而在中肠和血液组织cDNA的扩增产物中不存在。本研究对该丝腺特异的扩增小片段扩增产物进行了克隆鉴定,分析了其出现的原因,并探索了A3引物扩增的最佳扩增条件。

When studying genes related to cocoon color of Bombyx mori, we surprisingly discovered a small non-specific band in the silk gland. While using RT-PCR to amplify and detect with the A3 primers, we noticed that this fragment was unique and stable in the silk gland but it did not exist in the midgut and blood. Thus, we cloned the specific fragment and made bioinformatics study to analyze the possible reasons for its occurrence.