| 摘要: |
| 稀有鱼句鲫是一种新型的模式实验鱼，具有利用转基因技术培育开发成为观赏性鱼类的潜在可能。本实验采用PCR方法，从稀有鱼句鲫基因组DNA中分离得到大小为1 584bp的β肌动蛋白（β-actin）基因片段。序列分析表明，该片段包括长为1 507bp的启动调控区和77bp的部分转录区序列。启动调控区包括一段长为109bp β-actin基因上游调控序列，不翻译的外显子1和内含子1。上游调控序列中含有TATA box，CAAT box和CArG box等与转录活性密切相关的作用元件,并且在稀有鱼句鲫β actin基因第一个内含子中也含有1个CArG调控元件。将该启动子片段克隆到绿色荧光表达载体（pAcGFP1-1）上，显微注射入稀有鱼句鲫的受精卵中，通过荧光显微镜能观察到绿色荧光蛋白的表达。本实验成功分离了具有驱动活性的稀有鱼句鲫β-actin基因启动子。 |
| 关键词: 稀有鱼句鲫 β-肌动蛋白 启动子 序列分析 转基因 绿色荧光蛋白 |
| DOI： |
| 分类号: |
| 基金项目:Gobiocypris rarusβ actinPromoterSequence analysis |
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| Cloning of β-actin promoter of Gobiocypris rarus and detection of its transcription activity |
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| Abstract: |
| As a new laboratory fish in China, Gobiocypris rarus has the potential to develop into an ornamental fish by use of the transgenic techniques.A DNA fragment of 1584bp of β-actin gene from G.rarus genome was obtained by PCR,which included a 1507bp of a regulatory region and a 77bp of partial open reading frame (ORF).The regulatory region included a 109bp of 5′proximal promoter,an untranslated exon,and an intron of β-actin gene.The proximal promoter region contained consensus sequences of TATA box, CCAAT box and CArG box which were critical for transcription activity. Moreover, the intron also contained a typical CArG box.This promoter region fragment was inserted into green fluorescent protein gene vector (pAcGFP1-1),and the recombinant is then injected into the fertilized eggs of G. rarus by the method of micro-injection.The green fluorescence was observed under micro fluoroscope. This study proves that the cloned β-actin promoter has effective transcription activity. |
| Key words: Gobiocypris rarus β-actin promoter Sequence analysis Transgene green fluorescent protein |
