Development of transposon-vector-systems in insects.

昆虫转座子载体系统的开发。

• 批准号: 07554067
• 负责人: NITASAKA Eiji
• 金额: $ 2.3万
• 依托单位: KYUSYU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07554067/
• 关键词: Transposon; Drosophila; Bombyx mori; Mosquito; Vcctor; Transformation; 力

Transformation techniques are most important to analyze gene functions in model organisms, e. g. a mutant is transformed with a wild-type gene or a modified one. In insects, effective method for transformation had not been developed except for Drosophila. Using a transposable element, P-element, Drosophila can be transformed with high efficiency. Another advantage of this method is that tha most transgene is inserted with single copy and it eases to analyze given gene functions.The aim of this project is to develop transformation system using transposable elements. The materials used in this study are : Drosophila melanogaster, a mosquito species (Aedes albopictus) and the silkworm (Bombyx mori). Some species of mosquito are known as vectors for diseases, e. g. malaria, dengue. The silk worm is a useful insect not only for the textiles but also for the production of biomolecules from fibroin-gland. Therefore, an effetive transformation systems were desired in these species.Electroporat … More ion method is now widely used to transform organisms from bacteria to mammals. We tried to aply this method to insects using Drosophila embryos. In preliminary experiment with a cloned alpha-amylase gene as a reporter gene, the amylase activity was detected in the third instar larvae. Using P-element vector and a helper, however, no transformant was obtained out of 3000 embryos.To apply an eye color gene as a transformation marker, eye color mutations of Aedes albopictus were induced by Ethyl methan sulfonate (EMS). Until now, two mutations were identified. One is brown eye1 located on the sex-chromosome and anther is brown eye2 located on an autosome.Bm1 is a kind of SINE element of Bombyx mori. We had determined the complete sequences of fifteen of complete Bm1.1 elements. Sequence comparisons and a phylogenetic tree showed that they are classified into three groups. These results suggest that Bm1 was originated from tree ancestral elements and dispersed in the genome by reverse-transcription. Less

转化技术是分析模式生物基因功能的重要手段。G.用野生型基因或修饰的基因转化突变体。在昆虫中，除果蝇外，尚未开发出有效的转化方法。利用转座因子P-元件可以高效地转化果蝇。该方法的另一个优点是大多数转基因都是以单拷贝插入的，便于分析给定基因的功能。本研究中使用的材料是：黑腹果蝇、一种蚊子（白纹伊蚊）和家蚕（家蚕）。有些种类的蚊子被认为是疾病的传播媒介，例如：G.疟疾登革热家蚕是一种重要的昆虫，不仅可用于纺织，而且可用于从丝素腺中生产生物分子。因此，需要一种有效的转化系统在这些物种中。 ...更多信息 离子法现在被广泛用于将生物体从细菌转化为哺乳动物。我们尝试用果蝇胚胎将这种方法应用于昆虫。以已克隆的α-淀粉酶基因为报告基因，对3龄幼虫进行了淀粉酶活性检测。为了将眼色基因作为转化标记，用甲基磺酸乙酯（EMS）诱导白纹伊蚊（Aedes albopictus）眼色突变。到目前为止，已经发现了两种突变。Bm 1是家蚕的一种SINE元件，其主要功能是在家蚕的性染色体和常染色体上分别定位一个棕色眼1和棕色眼2。我们已经确定了15个完整的Bm1.1元素的完整序列。序列比较和系统发育树显示它们可分为三组。这些结果表明，Bm 1基因起源于树祖先元件，并通过逆转录分散在基因组中。少

项目成果

Yukuhiro,K: "Shared differences in Bm1.1 copies and their similarity to the Drosophila ARG-tRNA sequence." Japanese Journal of Genetics. 70. 748 (1995)

Yukuhiro,K：“Bm1.1 拷贝的共同差异及其与果蝇 ARG-tRNA 序列的相似性。”

Nitasaka, E.: "The molecular analysis of brown eye color mutations isolated from geographically discrete populations of Drosphila melanogaster." Molecular and General Genetics. 247. 164-168 (1995)

Nitasaka，E.：“对从地理上离散的黑腹果蝇种群中分离出的棕色眼睛颜色突变进行分子分析。”

Nitasaka, E.: "The molecular analysis of brown eye color mutations isolated from geographically discrete populations of Drosphila melanogaster." Molecular & General Genetics. 247. 164-168 (1995)

Nitasaka，E.：“对从地理上离散的黑腹果蝇种群中分离出的棕色眼睛颜色突变进行分子分析。”

Nagaraju, J: "Attempt at transgenesis of the silkworm(Bombyx mori L.)by egg-injection of foreign DNA." Appl.Entomol.Zool. 31. 587-596 (1996)

Nagaraju, J：“尝试通过卵子注射外源 DNA 来转基因蚕（Bombyx mori L.）。”

Sezutsu, H.: "Evolution of the LINE-like retrotransposon I element in the Drosophila melanogaster species subgroup." Mol. Gen. Gen.249. 168-178 (1995)

Sezutsu, H.：“果蝇物种亚群中 LINE 样逆转录转座子 I 元件的进化。”