Isolation and characterization of the Drosophila ornithine decarboxylase locus: evidence for the presence of two transcribed ODC genes in the Drosophila genome.

Isolation and characterization of the Drosophila ornithine decarboxylase locus: evidence for the presence of two transcribed ODC genes in the Drosophila genome.
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果蝇鸟氨酸脱羧酶基因座的分离和表征:果蝇基因组中存在两个转录 ODC 基因的证据。

DOI:
10.1089/dna.1993.12.499
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发表时间:
1993
影响因子:
3.1
通讯作者:
Chaim
Chaim
中科院分区:
生物学4区
文献类型:
--
作者:
E. Rom;Chaim

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采用聚合酶链反应(PCR)技术克隆了果蝇鸟氨酸脱羧酶(ODC)基因。在以cDNA或基因组DNA为模板的PCR中,使用两种简并寡核苷酸的混合物作为相对引物,所述简并寡核苷酸对应于在来自广泛分歧的物种的ODC之间完全保守的肽。所得DNA产物的序列分析证实其为ODC片段。然后将基因组PCR产物用作用于筛选果蝇基因组文库的探针,导致分离代表两个不同ODC基因(dODC 1和dODC 2)的基因组克隆。两个基因的序列分析表明,虽然在其编码区和非编码区不同,但它们的整体结构非常相似,包含6个外显子和5个短内含子。Southern杂交和序列分析表明,这两个ODC基因是以一个串联的头到尾的配置。两个ODC基因通过原位杂交分析被分配到第二染色体右臂上的位置44A。对应于这两个ODC基因的cDNA克隆的分离表明两者都在成蝇中转录。我们希望果蝇ODC基因组和cDNA克隆的分离将允许ODC在果蝇发育过程中的表达和多胺在这个过程中的作用的调查。
The polymerase chain reaction (PCR) was used to isolate two Drosophila ornithine decarboxylase (ODC) genes. Two mixtures of degenerate oligonucleotides corresponding to peptides that are fully conserved among ODCs from widely diverged species were used as opposing primers in the PCR with cDNA or genomic DNA as templates. Sequence analysis of the resulting DNA products confirmed their identity as ODC fragments. The genomic PCR product was then used as a probe for screening a Drosophila genomic library, resulting in the isolation of genomic clones representing two distinct ODC genes (dODC1 and dODC2). Sequence analysis of both genes demonstrated that although varying at their coding and noncoding regions, their overall structure is extremely similar containing 6 exons and 5 short introns. Southern blot and sequence analyses revealed that the two ODC genes are arranged in a tandem head-to-tail configuration. Both ODC genes were assigned by in situ hybridization analysis to position 44A on the right arm of the second chromosome. The isolation of cDNA clones corresponding to these two ODC genes demonstrated that both are transcribed in the adult fly. We hope that the isolation of genomic and cDNA clones of Drosophila ODC will permit the investigation of the expression of ODC during Drosophila development and the role of polyamines in this process.
DOI: 10.1126/science.2928784
发表时间: 1989-03-17
期刊: SCIENCE
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通讯作者: COFFINO, P
DOI: --
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使用 DNA 聚合酶链式反应进行同源性探测:从多个物种中分离编码琥珀酸脱氢酶铁硫蛋白的部分 cDNA 或基因组克隆。
DOI: 10.1073/pnas.86.6.1934
发表时间: 1989
影响因子: 11.1
作者:
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影响因子: 10.5
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