| 三角帆蚌Nacrein基因克隆、蛋白提纯及其对珍珠晶体成型的影响 |
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| 引用本文： | 韩健,李文娟,施志仪,郝莹莹,靳雨丽.三角帆蚌Nacrein基因克隆、蛋白提纯及其对珍珠晶体成型的影响[J].生物技术通报,2010(12). |
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| 作者姓名： | 韩健 李文娟 施志仪 郝莹莹 靳雨丽 |
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| 作者单位： | 上海海洋大学,农业部水产种质资源与养殖生态重点开放实验室,上海,201306 |
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| 基金项目： | 上海市科委基础重大项目(06aj14003); 上海市博士后科研资助计划面上项目(09R21413200); 上海市重点学科水生生物学建设项目(S3070) |
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| 摘 要： | 为研究三角帆蚌Nacrein蛋白对珍珠晶体成型的影响,通过巢式及3′-RACE PCR对三角帆蚌基质蛋白Nacrein基因3′端序列进行克隆,得到850bp碱基片段,分析表明其与合浦珠母贝同源基因序列相似度为56%,与马氏珠母贝相似度为50%。试验通过水提法初步分离出珍珠质水溶性基质蛋白,并采用快速蛋白液相色谱(fast protein liquid chromatography,FPLC)分离出分子量约为60kD的Nacrein蛋白。此外,采用配对试验设计,对试验组外套膜组织培养样品加入Nacrein蛋白,研究Nacrein蛋白对晶体成型的影响,结果显示,加入Nacrein蛋白的试验组结晶成棱形状;而未加Nacrein蛋白的对照组结晶成雪花状,而且在晶体形成的时间上,试验组也较之对照组要早。研究表明,Nacrein蛋白能加速外套膜组织分泌的钙质形成棱状结晶,从而对晶体成型产生影响。本研究为进一步研究基质蛋白对生物矿化的作用提供试验依据,对珍珠培育生产有一定指导意义。
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| 关 键 词： | Nacrein基因克隆 Nacrein基质蛋白 珍珠 晶体成型 |
Nacrein Gene Clone,Protein Extraction and Its Effect on Crystal Growth in Hyriopsis cumingii Lea |
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| Han Jian,Li Wenjuan,Shi Zhiyi,Hao Yingying,Jin Yuli.Nacrein Gene Clone,Protein Extraction and Its Effect on Crystal Growth in Hyriopsis cumingii Lea[J].Biotechnology Bulletin,2010(12). |
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| Authors: | Han Jian Li Wenjuan Shi Zhiyi Hao Yingying Jin Yuli |
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| Institution: | Han Jian Li Wenjuan Shi Zhiyi Hao Yingying Jin Yuli(Key Laboratory of Aquatic Genetic Researches and Aquacultural Ecology Certificated by the Ministry of Agriculture,Shanghai Ocean University,Shanghai 201306) |
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| Abstract: | In order to probe the effect of crystal growth on nacrein protein,the nest PCR and 3'-RACE PCR were used to clone Nacrein gene of Hyriopsis cumingii Lea.The target sequence(about 850 bp)was obtained.The Sequence were analyzed using DNAMAN software,and results showed that it had 56%and 50% similarity compared to homologous gene in Pinctada fucata and Pinctada martensii.The Nacrein protein was isolated and purified by the FPLC(fast protein liquid chromatography).The molecular weight of Nacrein protein was abo... |
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| Keywords: | Nacrein gene clone Nacrein protein extract Pearl Crystal growth |
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