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用于果蝇中P因子介导的基因转移的载体。

Vectors for P element-mediated gene transfer in Drosophila.

作者信息

Rubin G M, Spradling A C

出版信息

Nucleic Acids Res. 1983 Sep 24;11(18):6341-51. doi: 10.1093/nar/11.18.6341.

Abstract

We have constructed and tested several new vectors for P element-mediated gene transfer. These vectors contain restriction sites for cloning a wide variety of DNA fragments within a small, non-autonomous P element and can be used to efficiently transduce microinjected DNA sequences into the germ line chromosomes of D. melanogaster. The P element in one vector also carries the rosy gene which serves as an easily scored marker to facilitate the transfer of DNA fragments that do not themselves confer a recognizable phenotype. The failure of certain P element constructs to function as vectors suggests that P element sequences, in addition to the 31 bp inverse terminal repeats, are required in cis for transposition. Moreover, removal of the first 38 bp of the autonomous 2.9 kb P element appears to destroy its ability to provide a trans-acting factor (s) required for the transposition of non-autonomous P elements. Finally, we describe a genomic sequence arrangement that apparently arose by the transposition of a 54 kb composite P element from a tetramer plasmid.

摘要

我们构建并测试了几种用于P因子介导的基因转移的新载体。这些载体在一个小的非自主P因子内含有用于克隆多种DNA片段的限制性酶切位点,可用于有效地将显微注射的DNA序列转导到黑腹果蝇的种系染色体中。其中一个载体中的P因子还携带了朱红眼基因,该基因作为一个易于评分的标记,便于转移那些本身不赋予可识别表型的DNA片段。某些P因子构建体不能作为载体发挥作用,这表明除了31 bp的反向末端重复序列外,P因子序列顺式作用对于转座是必需的。此外,去除自主的2.9 kb P因子的前38 bp似乎会破坏其提供非自主P因子转座所需反式作用因子的能力。最后,我们描述了一种基因组序列排列,它显然是由一个54 kb复合P因子从四聚体质粒中转座产生的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a553/326377/34cfb43349d4/nar00363-0200-a.jpg

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