P Element-Mediated Transformation with the rosy Gene in Drosophila melanogaster
P Element-Mediated Transformation with the rosy Gene in Drosophila melanogaster
复制标题
P元件介导的黑腹果蝇rosy基因转化
DOI：
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
M. G. Kidwell
中科院分区：
文献类型：
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作者：
Wook Kim;M. G. Kidwell
我们用两种P元件构建体Pc[（ry+）B]和p[（ry+）SX 9]进行了D.黑腹菌将在自主P元件内携带rosy基因的Pc[（ry+）B]构建体注射到携带ry 506的真M菌株中。突变用于显微注射和转化的转座酶来源由命名为p-2- 3 hs的P元件辅助质粒提供，其与非自主P[（ry+）SX 9]构建体共注射到相同的ry 506 M菌株中。采用脱角法，从1143个G 0注射中获得了35个独立转化系（35/1143）。大约20%的注射胚胎作为成年人孵化。在G 0羽化苍蝇，约40%表现出眼睛的颜色是类似于野生型（ry+），但约60%的肥沃的G 0转化系似乎没有G1转化体。因此，G 0表达不太可能需要rosy转座子整合到染色体中。发现Pc[（ry+）B]和P[（ry+）SX 9]构建体在元件介导的转化频率上几乎相同。在此基础上，非自主P元件构建体可作为P元件介导的转化载体，在昆虫群体中导入和固定基因。
We have used two kinds of P element constructs, Pc[(ry+)B] and p[(ry+)SX9], for genetic transformation by microinjection of D. melanogaster. Pc[(ry+)B] construct carrying the rosy gene within an autonomous P element was injected into a true M strain caring the ry506. mutation. The source of transposase for microinjection and transformation was provided by a P element helper plasmid designated p-2-3hs, which was co-injected with nonautonomous P[(ry+)SX9] construct into same ry506 M strains. A dechorination method was adopted and 35 independent transformed lines were obtained froin 1143 G0 Injected (35/1143). About 20% of the injected embryos eclosed as adults. Among G0 eclosed flies, approximately 40% exhibited eye color that was similar to wild-type (ry+), but about 60% of fertile G0 transformed lines appeared to have no G1 transformants. Therefore it is unlikely that G0 expression requires integration of the rosy transposon into chromosomes. Pc[(ry+)B] and P[(ry+)SX9] constructs were found to be nearly same in the frequency of element-mediated transformation. On the basis of these results, nonautonomous P elements constructs could he used as same effective vectors in P element-mediated transformation for introducing and fixing genes in insect populations.