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海胆肌动蛋白基因序列的组织:编码细胞质肌动蛋白的含内含子DNA序列的分子克隆

Organization of actin gene sequences in the sea urchin: molecular cloning of an intron-containing DNA sequence coding for a cytoplasmic actin.

作者信息

Durica D S, Schloss J A, Crain W R

出版信息

Proc Natl Acad Sci U S A. 1980 Oct;77(10):5683-7. doi: 10.1073/pnas.77.10.5683.

Abstract

Southern transfer and solution hybridization experiments, using as probe a DNA fragment that encodes for Drosophila actin, demonstrate cross hybridization to DNA from the sea urchin Strongylocentrotus purpuratus. Recombinant DNA clones that contained sea urchin genomic DNA fragments were constructed and screened for the presence of actin-encoding DNA sequences by colony hybridization with the Drosophila actin sequence. Two different putative actin-encoding clones were identified and were shown to specifically hybridize actin-encoding mRNA from a complex mRNA population. Southern blot hybridization experiments with both the Drosophila actin sequence and one of the cloned sea urchin sequences, in conjunction with solution hybridization data, suggest an actin gene copy number of 5-20 per haploid genome. DNA sequence analysis of one of the cloned sequences indicates that this fragment codes for a cytoplasmic form of actin and contains an intervening sequence of at least 200 nucleotides beginning immediately after amino acid 121 in the protein sequence.

摘要

使用编码果蝇肌动蛋白的DNA片段作为探针进行Southern印迹转移和溶液杂交实验,结果表明该片段与紫球海胆(Strongylocentrotus purpuratus)的DNA发生了交叉杂交。构建了包含海胆基因组DNA片段的重组DNA克隆,并通过与果蝇肌动蛋白序列进行菌落杂交,筛选出含有肌动蛋白编码DNA序列的克隆。鉴定出两个不同的假定肌动蛋白编码克隆,并证明它们能与复杂mRNA群体中的肌动蛋白编码mRNA特异性杂交。用果蝇肌动蛋白序列和一个克隆的海胆序列进行Southern印迹杂交实验,并结合溶液杂交数据,表明单倍体基因组中肌动蛋白基因的拷贝数为5 - 20个。对其中一个克隆序列进行DNA序列分析表明,该片段编码一种细胞质形式的肌动蛋白,并且在蛋白质序列中氨基酸121之后紧接着含有一个至少200个核苷酸的间隔序列。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a609/350133/4ef027d69fc7/pnas00497-0145-a.jpg

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