CAJ | 학술논문

分离新的与B细胞活化相关基因。方法采用差异显示反转录PCR(DDRT-PCR)技术对人扁桃体活化和静止B细胞mRNA的差异表达进行分析，差异显示的片段经过Northern杂交验证后，作为探针进行人活化B细胞cDNA文库的筛选。结果差异显示分析共获得明显的差异表达的标签序列(expresed sequence tag,EST)62条，其中主要在静止B细胞表达的有32条，在活化B细胞表达的有30条。经Northern杂交验证，共获得阳性的片段25条。以在活化B细胞中高表达的EST30为探针，经3轮筛选人活化B细胞文库后获得1个新的全长为2 048 bp的cDNA克隆。该克隆含有1个630bp的开放读码框。其推断的氨基酸序列N端与酵母的动力蛋白KAR3部分同源。结论克隆了1条可能与B细胞活化相关的新基因。
분리신적여B세포활화상관기인。방법채용차이현시반전록PCR(DDRT-PCR)기술대인편도체활화화정지B세포mRNA적차이표체진행분석，차이현시적편단경과Northern잡교험증후，작위탐침진행인활화B세포cDNA문고적사선。결과차이현시분석공획득명현적차이표체적표첨서렬(expresed sequence tag,EST)62조，기중주요재정지B세포표체적유32조，재활화B세포표체적유30조。경Northern잡교험증，공획득양성적편단25조。이재활화B세포중고표체적EST30위탐침，경3륜사선인활화B세포문고후획득1개신적전장위2 048 bp적cDNA극륭。해극륭함유1개630bp적개방독마광。기추단적안기산서렬N단여효모적동력단백KAR3부분동원。결론극륭료1조가능여B세포활화상관적신기인。
Objective To clone the novel activation-related gene of B lymphocyte. Methods The differential display reversal transcription PCR (DDRT-PCR) technique was applied to analyse the expression difference of mRNA between resting and activated B lymphocyte from human tonsil. The positive differential display cDNA fragment identified by Northern-blotting was chosen as probe to filtrate human activated B lymphocyte cDNA library. Results Sixty two differential display cDNA fragments(expressed sequence tag, EST)were obtained. Thirty-two of them were mainly expressed in resting B lymphocyts and thirty were expressed in activated cells. Twenty-five were positive ones after identification by Northern blot analysis. A novel cDNA clone was obtained after using EST30 as a probe to filtrate the human activated B cell cDNA library.The whole cDNA clone was 2 048 bp in length and contains a 630 bp open reading frame. The N end of the deduced amino acid sequence was homologous with KAR3 protein which is a member of kinesins superfamily in yeast. Conclusions A novel possible activation-related gene in human B lymphocyte was obtained.