| 我国渔用疫苗的研制 |
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| 引用本文： | 杨先乐.我国渔用疫苗的研制[J].水产学报,2006,30(3):416-420. |
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| 作者姓名： | 杨先乐 |
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| 作者单位： | 上海水产大学生命科学与技术学院,上海,200090 |
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| 基金项目： | 农业部重点实验室基金;上海市学科建设项目 |
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| 摘 要： | 定量（quantitive PCR）是用PCR技术评估样本中靶基因的分子数，实现对核酸信息的量化分析及比较。目前主要有外标法和内标法两大类。其中，其中外标法中主要有PCR产物的直接定量法，极限稀释法等；内参标法中主要有竞争性和非竞争性PCR等。
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| 关 键 词： | 鲫 半定量PCR 内参物 雌激素样效应 雌激素受体 |
| 文章编号： | 1000-0615（2006）03-0416-05 |
| 收稿时间： | 2005-04-11 |
| 修稿时间： | 2005-04-11 |
The development of aquaculture vaccines in China |
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| YANG Xian-le.The development of aquaculture vaccines in China[J].Journal of Fisheries of China,2006,30(3):416-420. |
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| Authors: | YANG Xian-le |
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| Institution: | College of Aqua-life Science and Technology, Shanghai Fisheriea University, Shanghai 200090, China |
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| Abstract: | Semiquantitive PCR is a method to evaluate expression of target genes by analyzing RT-PCR products in comparison to standards.Two measures are involved;one is that primers for target gene and internal standard are put into the same tube to run PCR,and the other one is that primers for target gene and internal standard run PCR in separate tubes.In research of environmental endocrine disruption semiquantitive PCR is a usual means to measure estrogen-like effects which result from endocrine disruptors.In order to discuss advantages and disadvantages of these two methods for further research on endocrine disruption in Carassius auratus,RT-PCR was used to evaluate of expression of 776 bp of estrogen receptor gene(ER gene) in liver after taking the injection of E2 in C.auratus by following these two methods respectively,with 18S as internal standard.The results showed that ER gene and 18S gene were amplified successfully following both methods.The first method-putting primers for different gene in the same tube to run PCR might contain problems such as competition and different plat phase between target gene and internal standard,and it is needed to grope appropriate ratio of primers and numbers of cycles,yet it is more accurate.And another method putting primers for different genes in separate tubes to run PCR is easier because of the absence of no problemsexisted in the first method,whereas it is less accurate than the first one.In addition,based on the results,18S is an ideal internal standard for both methods in the research of estrogen-like effects in C.auratus. |
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| Keywords: | Carassius auratus semiquantitive PCR internal standard estrogen-like effects estrogen receptor(ER) |
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