分别采用Sepharose-6B凝胶过滤层析、Phenyl Sepharose FF疏水作用层析和rProtein A Sepharose亲和层析技术纯化黄鳍棘鲷(Acanthopagrus latus)血清IgM,并将这3种层析纯化方法进行了比较。纯化产物进行SDS-PAGE,电泳结果扫描后经Band Scan5.0软件分析显示,单独使用Sepharose-6B凝胶过滤层析或Phenyl Sepharose FF疏水作用层析得到的IgM纯度只有56%左右;二者联合使用纯度可以达到69%,而rProtein A Sepharose亲和层析一步就可以达到89%的纯度;纯化得到的黄鳍棘鲷血清IgM的重链和轻链分子量分别为73.6kD和26.5kD。本研究还以rProtein A Sepharose亲和层析纯化的黄鳍棘鲷血清IgM为抗原,制备其兔抗血清,采用WesternBlot和双向琼脂扩散检测抗血清免疫学活性,并用间接ELISA检测其效价达到1∶25600,为进一步开展黄鳍棘鲷免疫学方面的研究奠定了基础。
Scallops,comprise more than 300 species identified in worldwide oceans,occupy an economically important position in many coastal countries.Until now,more than 15 scallop species have been considered as valuable fishery or aquaculture resources.Scallop adductor muscles are being considered as one of the most flavorful sea foods in markets at home and abroad,mainly due to their delicious taste and abundant nutrition.Because of the potential values,the scallop aquaculture has made great progress during the past three decades in many countries,especially in China.For examples,the farmed tideland for scallop aquaculture has exceeded 1 million hectares(ha.) and the annual production is more than 11 million tons in 2007,which makes China the first-rank not only in farmed acreage but also in annual production.However,with the blooming expansion of scallop aquaculture,some problems such as poor growth of some wild strains and large-scale breakout of diseases have been arising during the past years.Further investigation and solution to these problems mainly depend on the availability of primary molecular genetic tools and the combination of efficient breeding programs with these molecular tools(i.e.molecular marker assisted selection,MAS).Microsatellite analysis based on polymerase chain reaction(PCR) offers the finest resolution for studying such genetic questions in aquacultural species.Based on the studies to date,microsatellite DNA marker has been one of the leading molecular markers for genetic analysis.Four scallops including Zhikong scallop Chlamys farreri,Noble scallop C.nobilis,Yesso scallop Patinopecten yessoensis and Bay scallop Argopecten irradians are the main species maricultured in China.In this study,we reviewed the progress in development of microsatellite DNA markers for these four scallops.We also summarized the applications of this robust molecular marker system in genetic analysis,such as genetic diversity analysis,pedigree reconstruction,genetic linkage map construction and species identification.
