The Study of DNA Replication-Related Proteins and Population Growth in Planktonic Algae

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簡歷

基本資料

Project title

Code/計畫編號

NSC88-2313-B019-044

Translated Name/計畫中文名

浮游藻類的核酸複製相關蛋白與族群生長研究

Project Coordinator/計畫主持人

Jeng Chang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=464822

Year

1999

Start date/計畫起

01-08-1998

Expected Completion/計畫迄

01-07-1999

Bugetid/研究經費

657千元

ResearchField/研究領域

生物科學
生物技術(理)

本計劃的長程目標在於辨識與浮游藻類細胞增殖有關的基因及蛋白質,以作為自然水域中族群生長的指標。在過去數年中,申請人已自微細藻類中得到兩種核酸複製所需的基因片段---「DNA 聚合脢 .alpha.」(DNA polymerase .alpha.) 及「增殖細胞核抗原」(proliferating cell nuclear anti-gen,PCNA)。在本計畫執行期間,主要的工作為以一種沿岸常見浮游矽藻(Skeleto-nema costatum,骨藻)為材料,偵測這兩種基因的表現,藉由這兩種基因產生的 mRNA與族群生長的關係,來評估何者較適宜作為海洋環境中矽藻族群生長的指標。實驗上以競爭性定量反轉錄聚合脢連鎖反應(competitive quantitative reverse transcription-polymerase chain reac-tion;簡稱CQ-RT-PCR)為手段,來偵測特定 mRNA 的含量。發現在各族群生長階段中,生長活躍的細胞比起平衡期的細胞含有較多的 DNA 聚合脢 .alpha. 和 PCNA 的mRNA。而在紫外線的處理實驗中,UVC 會損害骨藻的 DNA 並造成大量的T-dimer 的形成,這些細胞中 DNA 聚合脢 .alpha. 和 PCNA 的 mRNA 含量均較控制組的含量多。這顯示 DNA聚合脢 .alpha. 和 PCNA可能都參與 DNA 修補的工作。另一方面,在UVA/B 的處理中,PCNA 及 DNA 聚合脢 .alpha. 的mRNA 含量都與控制組類似。由於 UVA/B 是自然界中紫外線的主要形式,結果顯示使用 PCNA 或 DNA 聚合脢 .alpha.的 mRNA 作為骨藻細胞週期的 S 期指標,應不會將在進行修補 DNA 的細胞誤認為在生長的細胞。 The long term goal of this study is to identify cell cycle related genes and gene products which can be used as markers for phytoplankton population growth. In the past, we have successfully cloned 2 gene fragments belonging to DNA polymerase .alpha. and proliferating cell nuclear antigen (PCNA), respectively, and both are essential for DNA replication in unicellular algae. During the execution period of this project, major efforts were devoted to distinguish which gene is more suitable for the estimation of phytoplankton growth rate. A diatom, Skeletonema costatum, was used in this study, and the mRNA content of these 2 genes was detected by competitive quantitative reverse transcription-polymerase chain reaction (CQ-RT-PCR) at various growth stages and different UV-treatment conditions. During the exponential phase, population growth rate reached 0.6 day/sup -1/. The level of DNA polymerase .alpha. mRNA in exponential phase was higher than that found in the stationary phase. Similarly, PCNA mRNA level was higher during the exponential phase. In the UV treatment experiments, the DNA of S. costatum was severely damaged by UVC and a large quantity of thymidine-dimers was formed. The mRNA concentration of DNA polymerase .alpha. and PCNA were high in the UVC-treated samples. The results indicated that both DNA polymerase .alpha. and PCNA were involved in repairing DNA damages. In UVA/B treatment, T-dimer formation was less obvious, and mRNA concentrations of the 2 genes were similar to the controls. Since UVA/B is the major form of UV radiation in the natural environment, our results suggest that both the mRNA of DNA polymerase .alpha. and PCNA are suitable indicators for the estimation of population growth rate of S. costatum.

Keyword(s)

藻類
族群生長
核酸複製
蛋白質
浮游生物
Algae
Population growth
DNA replication
Protein
Plankton

DSpace CRIS

The Study of DNA Replication-Related Proteins and Population Growth in Planktonic Algae

基本資料

Description