CAJ | 학술논문

为了筛选链状亚历山大藻特异表达的发光相关基因,应用抑制消减杂交技术,构建了链状亚历山大藻特异表达的cDNA消减文库,共获得500个克隆,通过基因PCR初步筛选表明插入片段的大小主要集中在250～1000 bp,随机挑选10个阳性克隆进行双向测序和序列比对分析,有2个克隆基因片段与已知基因序列高度同源,分别为荧光素结合蛋白和羧化酶/加氧酶,另有8个克隆基因片段在GenBank中未查找到相应的同源基因,可能为未知新基因序列.这些基因的获得为进一步研究链状亚历山大藻特异表达基因,阐明其发光机理及建立特异甲藻的新型检测方法提供重要依据.
위료사선련상아력산대조특이표체적발광상관기인,응용억제소감잡교기술,구건료련상아력산대조특이표체적cDNA소감문고,공획득500개극륭,통과기인PCR초보사선표명삽입편단적대소주요집중재250～1000 bp,수궤도선10개양성극륭진행쌍향측서화서렬비대분석,유2개극륭기인편단여이지기인서렬고도동원,분별위형광소결합단백화최화매/가양매,령유8개극륭기인편단재GenBank중미사조도상응적동원기인,가능위미지신기인서렬.저사기인적획득위진일보연구련상아력산대조특이표체기인,천명기발광궤리급건립특이갑조적신형검측방법제공중요의거.
To isolate the bioluminescence specific expressed genes of dinoflagellate Alexandrium catenella(A. catenella), suppression subtractive hybridization was performed to construct the differential expressing cDNA libraries of Alexandrium catenella. A total of 500 cDNA clones were picked. PCR amplification revealed that the length of inserted fragments was mainly from 250 to 1000 base pairs. Ten randomly selected clones after PCR screening were sequenced and analysed in GenBank with Blast search. Two ESTs shared significant identity with the luciferin binding protein and Ribulose-1,5-bisphosphate carboxylase/oxygenase and the other ESTs represent novel genes of Alexandrium catenella with no significant homology in GenBank. These results had provided the foundation for cloning new specific genes of Alexandrium catenella and further studying and establishing the novel detection method for specific dinoflagellata.