安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
Acta Universitatis Medicinalis Anhui
2015年
9期
1207-1210
,共4页
钱成%王蓓华%徐雪琴%潘林鑫%李新颖%刘晓颖%范礼斌
錢成%王蓓華%徐雪琴%潘林鑫%李新穎%劉曉穎%範禮斌
전성%왕배화%서설금%반림흠%리신영%류효영%범례빈
DAG1%转染%基因表达%定位
DAG1%轉染%基因錶達%定位
DAG1%전염%기인표체%정위
DAG1%transfection%gene expression%localization
目的 研究肌营养不良蛋白( DG)及其衍生物α-DG、β-DG在真核细胞内的表达与定位. 方法 构建pcDNA3. 1-DAG1-FLAG、pcDNA3. 1-DAG1α-FLAG 和 pcDNA3. 1-DAG1β-FLAG 3个真核表达质粒,并分别转染至HEK 293T细胞中;提取总蛋白, Western blot检测表达情况;分别转染至COS7细胞,在荧光显微镜下观察细胞定位情况. 结果 成功构建了带有DAG1、DAG1α、DAG1β 基因的真核表达载体,且在HEK 293T细胞中都能有效表达,在COS7 细胞中主要分布于细胞质. 结论 DG 及其衍生物 α-DG、β-DG 在 HEK 293T、COS7细胞中均有表达,主要分布在细胞质内,为进一步了解DAG1的功能提供了细胞学基础.
目的 研究肌營養不良蛋白( DG)及其衍生物α-DG、β-DG在真覈細胞內的錶達與定位. 方法 構建pcDNA3. 1-DAG1-FLAG、pcDNA3. 1-DAG1α-FLAG 和 pcDNA3. 1-DAG1β-FLAG 3箇真覈錶達質粒,併分彆轉染至HEK 293T細胞中;提取總蛋白, Western blot檢測錶達情況;分彆轉染至COS7細胞,在熒光顯微鏡下觀察細胞定位情況. 結果 成功構建瞭帶有DAG1、DAG1α、DAG1β 基因的真覈錶達載體,且在HEK 293T細胞中都能有效錶達,在COS7 細胞中主要分佈于細胞質. 結論 DG 及其衍生物 α-DG、β-DG 在 HEK 293T、COS7細胞中均有錶達,主要分佈在細胞質內,為進一步瞭解DAG1的功能提供瞭細胞學基礎.
목적 연구기영양불량단백( DG)급기연생물α-DG、β-DG재진핵세포내적표체여정위. 방법 구건pcDNA3. 1-DAG1-FLAG、pcDNA3. 1-DAG1α-FLAG 화 pcDNA3. 1-DAG1β-FLAG 3개진핵표체질립,병분별전염지HEK 293T세포중;제취총단백, Western blot검측표체정황;분별전염지COS7세포,재형광현미경하관찰세포정위정황. 결과 성공구건료대유DAG1、DAG1α、DAG1β 기인적진핵표체재체,차재HEK 293T세포중도능유효표체,재COS7 세포중주요분포우세포질. 결론 DG 급기연생물 α-DG、β-DG 재 HEK 293T、COS7세포중균유표체,주요분포재세포질내,위진일보료해DAG1적공능제공료세포학기출.
Objective To investigate the expression and localization of DG and its derivative α-DG,β-DG in eu-karyotic cells. Methods To construct three eukaryotic expression plasmids pcDNA3. 1-DAG1-FLAG,pcDNA3. 1-DAG1α-FLAG and pcDNA 3. 1-DAG1β-FLAG. The plsamids were transfected into HEK 293T cells respectively and the expression was checked by Western blot. Their expressions in COS7 cells were detected by fluorescence mi-croscopy. Results The expression plasmids of pcDNA3. 1-DAG1-FLAG, pcDNA3. 1-DAG1α-FLAG and pcD-NA3. 1-DAG1β-FLAG were constructed successfully, which could be effectively expressed in HEK 293T and COS7 cells, and the localization of ( DG andα-DG,β-DG) mainly in the cytoplasm. Conclusion DG and its derivativeα-DG,β-DG can express efficiently in HEK 293T and COS7 cells, and all of them are mainly distributed in the cy-toplasm, which provide cellular basis to further understand the function of DAG1 gene.