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。
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 about 60 kD.Furthermore,the comparative test was designed（treatment group and control group）to study relationship between Nacrein protein and crystal growth.The total of 12 mantle tissues were cultured（6 dish/group）,and the tissues of treatment group were added the Nacrein protein in culture media,while there of control group had not added them.The research showed that Nacrein protein accelerated the growth of prismatic-shaped crystal by the acting on secretion of mantle tissue,for further research on biological mineralization matrix protein function to provide the foundations and certain significance on pearl development and production.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第12期137-141,共5页
Biotechnology Bulletin
基金
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)
