三角帆蚌(Hyriopsis cumingii)珍珠形成初期钙代谢的特征分析

采用生理生化及分子生物学方法,对刚植片的三角帆蚌进行了为期57天的研究,探讨三角帆蚌珍珠形成初期钙的代谢特征。实验每周取样一次,分别测定了珍珠囊和珍珠的直径,珍珠囊和珍珠的重量及水温,外套膜、鳃、围心腔三种组织的钙含量,酸、碱性磷酸酶的比活力,以及外套膜中钙调素基因的表达量。结果表明,植片后三角帆蚌的珍珠囊及珍珠在第22天和43天各出现一个快速生长期。其外套膜和鳃中的酸性磷酸酶比活力第8天升到最高,随后逐渐下降;外套膜中的碱性磷酸酶比活力第15天达到最高,第29天后开始快速下降;鳃和围心腔中的碱性磷酸酶比活力第15天后才开始上升,第29天达到最高。三种组织的钙含量总体呈现上升趋势,其中鳃的钙含量最高并在第43天达到最高值。植片后三角帆蚌外套膜中的钙调素RNA表达量明显增强,第8天和43天的表达量尤其显著。三角帆蚌珍珠的形成主要依赖于鳃和外套膜对钙的代谢,在此过程中酸、碱性磷酸酶以及钙调素起着重要的作用。可能依照钙调素基因上调表达、酸性和碱性磷酸酶激活、鳃和外套膜钙含量升高、珍珠生长的顺序来进行,植片后的第8天、15天和43天是珍珠形成的关键期。     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda (Bangiales, Rhodophyta)

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensiss,cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exous and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta）

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca²⁺ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.