黄鳝IRF-1和IRF-2保守序列的克隆与表达

Cloning and Expression Analysis of IRF-1 and IRF-2 in Rice-field Eel,Monopterus albus

利用简并引物扩增的黄鳝(Monopterus albus)IRF-1和IRF-2中间片段,大小分别为507bp和577bp。分别提取黄鳝性逆转前后的3个发育阶段(雌性、间性和雄性)脾脏和中肾组织的RNA,反转录成cDNA,分别利用IRF-1和IRF-2特异引物对这些cDNA模板进行PCR扩增,结果表明IRF-1和IRF-2在黄鳝3个不同的发育阶段表达量基本一致,没有明显的差异;为了进一步阐明IRF-1和IRF-2的表达规律,分别提取雄性黄鳝性腺、肌肉、血液、皮肤、肠、中肾、脾脏、肝脏、脑等9个组织的总RNA,反转录成cDNA,利用IRF-1和IRF-2特异引物对其进行PCR扩增,结果表明IRF-1和IRF-2在雄性黄鳝不同组织表达情况不同,IRF-1在血液、皮肤、肠、中肾和肝脏表达量较高,而IRF-2在中肾、肠和脾脏中表达量较高,IRF-1和IRF-2在黄鳝中均呈现组成型表达。

The partial cDNA of IRF-1 and IRF-2 were identified in rice-field eel(Monopterus albus),whose nucleotide was 507bp and 577bp respectively.The expression of IRF-1 and IRF-2 was observed respectively the female,the intersex and the male of rice-field eel,which showed the expression of four kind IRFs was not different in the three different upgrowth stages.In order to further clarify their spatial and temporal expression patterns,the total RNA of gonad,muscle,blood,skin,intestine,trunk kidney,spleen and brain of male rice-field were extracted respectively and then reversly transcribed into cDNA.The analysis of semi-quantitative RT-PCR revealed the expression pattern of the IRFs presented obvious discrepancies in the different organs.IRF-1 and IRF-2 were constitutively expressed in all organs that were examined.The IRF-1 expression levels was strong in the intestine,the trunk kidney,the liver,the skin and the blood while IRF-2 showed relative high expression in the trunk kidney,the intestine and the spleen.