NUMBER AND EVOLUTIONARY CONSERVATION OF ALPHA-TUBULIN AND BETA-TUBULIN AND CYTOPLASMIC BETA-ACTIN AND GAMMA-ACTIN GENES USING SPECIFIC CLONED CDNA PROBES
NUMBER AND EVOLUTIONARY CONSERVATION OF ALPHA-TUBULIN AND BETA-TUBULIN AND CYTOPLASMIC BETA-ACTIN AND GAMMA-ACTIN GENES USING SPECIFIC CLONED CDNA PROBES
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DOI：
10.1016/0092-8674(80)90238-x
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发表时间：
1980-01-01
期刊：
影响因子：
64.5
通讯作者：
KIRSCHNER, MW
中科院分区：
文献类型：
--
作者：
CLEVELAND, DW;LOPATA, MA;KIRSCHNER, MW
从胚胎鸡脑的mRNA构建含有对α-微管蛋白、β-微管蛋白、β-肌动蛋白和γ-肌动蛋白特异的插入DNA序列的细菌克隆。分离了分别含有大约75％、90％和>90％存在于α-微管蛋白、β-微管蛋白和β-肌动蛋白mRNA中的序列的质粒，以及含有β-和γ-肌动蛋白mRNA的广泛3''非翻译区的部分的克隆。 2 个微管蛋白的序列不交叉杂交。每个微管蛋白的标记克隆探针与用几种限制性核酸内切酶消化的鸡DNA的杂交揭示了大约4个α-微管蛋白片段和4个β-微管蛋白片段。这似乎是基因的数量，因为任一克隆的微管蛋白c[互补]DNA的5''和3''末端都与用限制性内切核酸酶消化的基因组DNA中的至少4个共同片段杂交。微管蛋白探针能够在严格条件下与所有测试的脊椎动物基因组的 DNA 以及海胆 DNA 杂交，但不能与酵母 DNA 杂交。在消化的海胆精子 DNA 中，有超过 20 个不同的片段与微管蛋白 cDNA 的 5'' 和 3'' 端杂交。全长 β-肌动蛋白 cDNA 克隆与限制性鸡 DNA 中的 4-7 个条带杂交，并与来自所有其他测试物种（包括海胆和酵母）的 DNA 交叉杂交。对β-和γ-肌动蛋白mRNA的3''非翻译区具有特异性的克隆探针与鸡DNA的杂交表明，β-肌动蛋白的3''非翻译区具有特异性。序列在基因组和γ中仅存在一次。最多存在 3 个副本。 3''非翻译序列在进化上都不是保守的。
Bacterial clones containing inserted DNA sequences specific for .alpha.-tubulin, .beta.-tubulin, .beta.-actin and .gamma.-actin were constructed from mRNA of embryonic chick brain. Plasmids containing approximately 75, 90 and > 90%, respectively, of the sequences present in .alpha.-tubulin, .beta.-tubulin and .beta.-actin mRNA were isolated, as were clones containing parts of the extensive 3'' untranslated regions of the .beta.- and .gamma.-actin mRNA. The sequences for the 2 tubulins do not cross hybridize. Hybridization of labeled, cloned probes for each of the tubulins with chicken DNA digested with several restriction endonucleases reveals about 4 fragments for .alpha.- and 4 for .beta.-tubulin. This seems to be the number of genes, since both the 5'' and 3'' ends of either cloned tubulin c[complementary]DNA hybridize to at least 4 common fragments in genomic DNA digested with restriction endonucleases. The tubulin probes are able to hybridize under stringent conditions to DNA of all vertebrate genomes tested and to sea urchin DNA, but not to yeast DNA. In digested sea urchin sperm DNA there are more than 20 different fragments which hybridize to both the 5'' and 3'' ends of the tubulin cDNA. A full-length .beta.-actin cDNA clone hybridizes to 4-7 bands in restricted chicken DNA and cross hybridizes to DNA from every other species tested, including sea urchin and yeast. Hybridization to chicken DNA of cloned probes specific for the 3'' untranslated regions of .beta.- and .gamma.-actin mRNA indicates that the .beta. sequence is present only once in the genome and the .gamma. is present in at most 3 copies. Neither 3'' untranslated sequence is conserved evolutionarily.