Molecular cloning of a novel sex pheromone responsible for the release of a different sex pheromone in Closterium peracerosum-strigosum-littorale complex.
Molecular cloning of a novel sex pheromone responsible for the release of a different sex pheromone in Closterium peracerosum-strigosum-littorale complex.
复制标题
一种新型性信息素的分子克隆，该性信息素负责在新月藻-strigosum-littorale复合体中释放不同的性信息素。
DOI：
10.1093/oxfordjournals.pcp.a029317
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发表时间：
1998
影响因子：
4.9
通讯作者：
Y. Kamiya
中科院分区：
文献类型：
--
作者：
H. Sekimoto;R. Fukumoto;N. Dohmae;K. Takio;T. Fujii;Y. Kamiya
一个性信息素，原生质体释放诱导蛋白（PR-IP）诱导剂，的新月藻peracerosum-strigosum-littorale复合体，是已知的诱导释放PR-IP，交配型加（mt+）细胞在有性生殖过程中。纯化的PR-IP诱导剂用胰蛋白酶处理以获得用于测定部分氨基酸序列的内部肽。使用这些序列，合成寡核苷酸并用作组合逆转录-PCR的引物。扩增296 bp的cDNA片段，允许克隆相应的全长cDNA（CpPI;新月藻-糙伏毛藻-滨海新月藻复合体PR-IP诱导物）。CpPI的推导的氨基酸序列编码M（r）23，071的212个氨基酸残基的蛋白质，而分泌的肽部分预测具有M（r）15，717的142个氨基酸残基，并且与已知蛋白质没有显着相似性。预测的蛋白质具有三个可能的天冬酰胺连接的糖基化位点的共有序列。当交配型负（mt-）细胞在光照下以低细胞密度孵育时，CpPI基因表达。培养基中的氮剥夺增强了CpPI基因的表达。基因组Southern杂交分析表明，cDNA探针杂交的几个DNA片段从两个基因组的MT-和MT+细胞。然而，在mt-细胞中，通过北方杂交不能检测到PR-IP诱导剂的转录物。
A sex pheromone, protoplast-release-inducing protein (PR-IP) inducer, of the Closterium peracerosum-strigosum-littorale complex is known to induce the release of PR-IP, from mating-type plus (mt+) cells during sexual reproduction. The purified PR-IP inducer was treated with trypsin to obtain internal peptides for determination of partial amino acid sequences. Using these sequences, oligonucleotides were synthesized and used as primers for the combined reverse transcription-PCR. A 296 bp cDNA fragment was amplified, permitting the cloning of corresponding full length cDNA (CpPI; Closterium peracerosum-strigosum-littorale complex PR-IP inducer). The deduced amino acid sequence of CpPI encodes a protein of 212 amino acid residues of M(r) 23,071 whereas portion of the peptide secreted is predicted to have 142 amino acid residues of M(r) 15,717 and shows no significant similarity with known proteins. The predicted protein has three possible consensus sequences for asparagine-linked glycosylation site. The CpPI gene was expressed when mating-type minus (mt-) cells were incubated at a low cell density in the light. Nitrogen deprivation from the medium enhances expression of the CpPI gene. An analysis by genomic Southern hybridization revealed that the cDNA probe hybridized to several DNA fragments obtained from both the genome of mt- and mt+ cells. However, in mt- cells, transcripts for the PR-IP inducer could not be detected by Northern hybridization.