The Production of an Antiserum against DNA Polymerase Alpha and Its Application to the Measurement of Phytoplankton Growth Rate in situ

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Project title

Code/計畫編號

NSC83-0209-B019-004

Translated Name/計畫中文名

製造能辨識核酸聚合脢之抗體來研究浮游植物在自然界的生長率

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=99808

Year

1994

Start date/計畫起

01-02-1994

Expected Completion/計畫迄

01-07-1994

Bugetid/研究經費

444千元

ResearchField/研究領域

生物科學

本計畫的目標是以聚合？連鎖反應(PCR)的技術在浮游藻類各主要分類群中尋找DNA聚合？.alpha.的同源基因(Homologous gene),以期在未來能針對這個蛋白質製作抗體並以此判定浮游藻類在自然水域中生長的快慢。計畫進行的頭半年(民國83年2月至7月)中,本實驗室之主要進展為利用PCR在單細胞藻類中增殖Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase)的基因片段,以求得針對單細胞藻類最恰當的反應條件。結果顯示PCR能在Skeletonema costatum(矽藻綱)及Tetraselmis chui(Prasinophyceae)中增殖出DNA片段,但以S. costatum的表現較為穩定,所得到的兩個DNA片段長度分別為400及450bp上下,均符合預期的大小。在未來的實驗中,類似的反應條件將被用來增殖S. costatum中的DNA聚合？.alpha.的基因片段,而且Rubisco的一對起動引子(Primers)可充作正控制組(Positive control)。 The goal of this ongoing study is to amplify phytoplankton gene fragments homologous to DNA polymerase .alpha. using the polymerase chain reaction technique (PCR). With such gene fragments in hands, we are hoping to produce antisera against phytoplankton DNA polymerase .alpha., and use the antisera as a convenient was to assess phytoplankton growth state in natural environments. During the first half year of this project, we chose to amplify a fragment of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) gene in order to determine a set of reaction conditions which is suitable for phytoplankton. We obtained positive results from 2 algal species, Skeletonema costatum (Bacillariophyceae) and Tetraselmis chui (Prasinophyceae). The performance of S. costatum was found to be more stable. The lengths of the 2 DNA fragments from S. costatum were about 400 and 450bp, respectively, and they agreed well with the anticipated sizes. In the future, similar conditions will be used to amplify DNA polymerase .alpha. gene fragments from S. costatum with Rubisco primers as the positive control.

Keyword(s)

浮游植物
DNA聚合酶
抗體
生長速率
聚合酶連鎖反應
Phytoplankton
DNA polymerase
Antibody
Growth rate
Polymerase chain reaction (PCR)

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The Production of an Antiserum against DNA Polymerase Alpha and Its Application to the Measurement of Phytoplankton Growth Rate in situ

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