| HE Shanying,YU Zhigang. 2008. Estimation of growth rate of Skeletonema costatum based on relative expression amount of PCNA gene. Acta Oceanologica Sinica, (6):149－158 |
| Estimation of growth rate of Skeletonema costatum based on relative expression amount of PCNA gene |
| Estimation of growth rate of Skeletonema costatum based on relative expression amount of PCNA gene |
| Received:July 21, 2008 Revised:November 10, 2008 |
| DOI： |
| Key words:Skeletonema costatum fluorescent quantitative PCR PCNA growth rate |
| 中文关键词: Skeletonema costatum fluorescent quantitative PCR PCNA growth rate |
| 基金项目:The Zhejiang Provincial Natural Science Foundation of China under contract No. Y5080149; the National Natural Scientifie Foundation of China under contract No. 40406028. |
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| Abstract: |
| Partial sequence of Skeletonema costatum proliferating cell nuclear antigen (PCNA) gene was obtained by reverse transcriptase PCR(RT-PCR) and 3' rapid amplification of cDNA ends (3'-RACE) techniques. Based on the obtained PCNA and cytochrome b gene(Cyt b gene) sequences, a real-time fluorescent quantitative PCR (FQ-PCR) method was developed to detect the expression S. costatum PCNA gene, and this method was applied to study the relationship between the growth rate of S. costatum and the average expression amount of PCNA gene in a single cell. The expression amount of PCNA gene had large variation in cells collected at different culture phases, and the trend was well consistent with the growth rate, which suggested that the expression amount of PCNA gene correlated well with the cell division, and the PCNA could be a promising indicator for the S. costatum cell proliferation. Furthermore, using the PCNA gene as the objective gene and the Cyt b gene as the house-keeping gene, a new method for estimating the in situ growth rate of S. costatum was established by analysis of the relative expression quantity (REQ) of the PCNA gene. |
| 中文摘要: |
| Partial sequence of Skeletonema costatum proliferating cell nuclear antigen (PCNA) gene was obtained by reverse transcriptase PCR(RT-PCR) and 3' rapid amplification of cDNA ends (3'-RACE) techniques. Based on the obtained PCNA and cytochrome b gene(Cyt b gene) sequences, a real-time fluorescent quantitative PCR (FQ-PCR) method was developed to detect the expression S. costatum PCNA gene, and this method was applied to study the relationship between the growth rate of S. costatum and the average expression amount of PCNA gene in a single cell. The expression amount of PCNA gene had large variation in cells collected at different culture phases, and the trend was well consistent with the growth rate, which suggested that the expression amount of PCNA gene correlated well with the cell division, and the PCNA could be a promising indicator for the S. costatum cell proliferation. Furthermore, using the PCNA gene as the objective gene and the Cyt b gene as the house-keeping gene, a new method for estimating the in situ growth rate of S. costatum was established by analysis of the relative expression quantity (REQ) of the PCNA gene. |
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