鳞头犬牙南极鱼zona pellucida蛋白抗冻功能的研究
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- 英文论文题名：Cloning of zona pellucida protein from Antarctic notothenioid fish Dissostichus mawsoni and identification of its cold tolerance in CHO cell
- 论文作者：付艳霞 ; 陈良标
- 英文论文作者：FU Yanxia ; Chen Liangbiao ; College of Fisheries and Life Sciences ; Key Laboratory of Aquaculture Resources and Utilization ; Ministry of Education ; Shanghai Ocean University
- 年：2016
- 作者机构：上海海洋大学水产与生命学院水产种质资源发掘与利用省部共建教育部重点实验室;
- 论文关键词：zona pellucida ; 南极鱼 ; 低温 ; 存活率
- 英文论文关键词：zona pellucida ; Antarctic notothenioid fish ; low temperature ; survival
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：562k
- 原文格式：D
- 会议级别：全国
摘要
鳞头犬牙南极鱼卵壳蛋白(zona pellucida,ZP)在长期低温环境下肽链长度发生了缩短的进化趋势,为探究肽链缩短所发生的生物学意义,本实验研究了ZP蛋白对于中国仓鼠卵巢细胞(Chinese hamster ovary,CHO)在低温下的存活率,从南极鱼卵巢中提取RNA并反转成cDNA克隆到表达载体PIRES2-EGFP,共构建了三种ZPC5质粒,并将质粒分别转染至CHO细胞中进行表达,低温处理后,采用流式细胞术对细胞的存活率进行测定。结果表明,鳞头犬牙南极鱼ZPC5基因在中国仓鼠卵巢细胞中高效表达,低温处理后,肽链长度较短的ZPC5基因的CHO细胞存活率越高,与未经低温处理的CHO细胞相比,目的蛋白表达量也明显增多。研究表明,南极鱼在进化过程中,一些鱼类的ZPC5基因在低温下具有提高细胞抵抗低温的功能,且在进化过程中发生了正选择。
In order to understand Dissostichus mawsoni of Antarctic fish zona pellucida protein peptide chain length in the process of evolution to shorten the evolution trend of select role, the experiment research the ZP protein for Chinese hamster ovary cells the rate of survival in low temperature, from Antarctic fish ovary extraction RNA and reverse into cDNA clone to expression vector PIRES2-EGFP, totally has three ZPC5 genes, and respectively transfected into CHO for expression, after processing, By flow cytometry for cell survival rates were determined. Results showed that the Antarctic D.mawsoni ZPC5 gene expression in CHO cells, cryogenic treatment,peptide chain length shortest ZPC5 gene in CHO cell survival rate is the highest, compared with without low temperature treatment of CHO cells, protein expression was also increased considerably. Studies showed that Antarctic fish during the course of evolution, ZPC5 gene has the function of cells that are more resistant to low temperature, and evolution has occurred positive selection.
In order to understand Dissostichus mawsoni of Antarctic fish zona pellucida protein peptide chain length in the process of evolution to shorten the evolution trend of select role, the experiment research the ZP protein for Chinese hamster ovary cells the rate of survival in low temperature, from Antarctic fish ovary extraction RNA and reverse into cDNA clone to expression vector PIRES2-EGFP, totally has three ZPC5 genes, and respectively transfected into CHO for expression, after processing, By flow cytometry for cell survival rates were determined. Results showed that the Antarctic D.mawsoni ZPC5 gene expression in CHO cells, cryogenic treatment,peptide chain length shortest ZPC5 gene in CHO cell survival rate is the highest, compared with without low temperature treatment of CHO cells, protein expression was also increased considerably. Studies showed that Antarctic fish during the course of evolution, ZPC5 gene has the function of cells that are more resistant to low temperature, and evolution has occurred positive selection.
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