Organization of actin gene sequences in the sea urchin: molecular cloning of an intron-containing DNA sequence coding for a cytoplasmic actin.
Organization of actin gene sequences in the sea urchin: molecular cloning of an intron-containing DNA sequence coding for a cytoplasmic actin.
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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。
DOI：
10.1073/pnas.77.10.5683
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发表时间：
1980
影响因子：
11.1
通讯作者：
CrainJr,WR
中科院分区：
文献类型：
--
作者：
Durica,DS;Schloss,JA;CrainJr,WR
以果蝇肌动蛋白编码的DNA片段为探针，进行了南方转移和溶液杂交实验，证明了与海胆（strongylocentrus purpuratus） DNA的交叉杂交。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的集落杂交筛选了肌动蛋白编码DNA序列的存在。鉴定了两个不同的假定的肌动蛋白编码克隆，并证明它们特异性地杂交了来自复杂mRNA群体的肌动蛋白编码mRNA。与果蝇肌动蛋白序列和一个克隆海胆序列的Southern blot杂交实验，结合溶液杂交数据，表明每个单倍体基因组的肌动蛋白基因拷贝数为5-20。对其中一个克隆序列的DNA序列分析表明，该片段编码肌动蛋白的细胞质形式，并且包含至少200个核苷酸的中间序列，该中间序列在蛋白质序列的第121个氨基酸之后立即开始。
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.