三角帆蚌Nacrein基因克隆、蛋白提纯及对珍珠晶体成型的影响.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 12 Nacrein 韩健 李文娟 施志仪 郝莹莹 靳雨丽 ( , 201306) : 为研究三角帆蚌 Nacrein蛋白对珍珠晶体成型的影 , 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 850 bp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 li u id chromatography, FPLC)分离出分子量约为 60 kD 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N acrein蛋白对晶体成型的影 , 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, N acrein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 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 : Nacrein基因克隆 Nacrein基质蛋白 珍珠 晶体成型 Nacrein Gene Clone, Protein Extraction and ItsEffect on CrystalGrowth inHyriop sis cum ing ii Lea H an Jian LiW enjuan Shi Zhiyi H ao Y ingy ing Jin Yuli (K ey Laboratory of A quatic Genetic Researches and Aquacultural Eco logy Certfi icated by theM inistry of Agriculture, ShanghaiOcean Uni ersity, Shanghai 201306) Abstract: In order to probe the effect of crystal grow th on nacrein protein, the nest PCR and 3 RACE PCR w ere used to clone Nacrein gene ofHy riop sis cum ing ii Lea. The target se uence ( about 850 bp) w as obtained. The Se uence were analyzed using DNA MAN softw are, and results show ed that it had 56% and 50% smi ilarity compared to hom ologous gene inP inctadaf uca ta andP inctada m artensii. The Nacrein protein was isolated and purified by the FPLC ( fast protein li u id chrom atography). The m olecular w eigh t of Nacrein protein w as about 60 kD. Furtherm ore, the com parative test w as designed ( treatm ent group and control group) to study rela tionship between Nacrein protein and crystal grow th. The total of 12 m antle tissu