摘要
目的:探讨InterAluPCR技术在筛选鉴定含人DNA小鼠转染细胞和分离人源DNA片段中的应用。方法:设计人特异性Alu引物,通过InterAluPCR扩增对转染人基因组DNA后回复突变的小鼠细胞进行鉴定筛选;分离插入的人源DNA片段;经原位PCR鉴定其人源性。结果:应用该技术从8株回复突变株中筛选出6株含特异性人DNA;从其中12号小鼠细胞基因组DNA中分离出3条人源DNA片段;PCR原位杂交确证所分离的DNA片段具有人的特异性。结论:InterAluPCR技术是应用于筛选鉴定含人DNA杂种细胞、分离其中人源插入片段的简便、有效的方法。
Objective: To identify the mouse tumor cell clones transfected with normal human genomic DNAs and to isolate the inserted human anonymous sequences by Inter Alu PCR. Methods: Human DNA fragments were amplified by Inter Alu PCR technique from normal human genomic DNAtransfected mouse tumor cell clones, of which the metastatic phenotype had been suppressed. The human origin of the amplified DNA was further confirmed by PCR in situ hybridization. Results: From eight revertants exhibiting normal phenotype, six clones containing human DNA were identified by Inter Alu PCR ; a continuum of human genomic DNA fragments was obtained from number 12 mouse tumor cell, of which the metastatic phenotype was suppressed. Conclusion: The results indicated that Inter Alu PCR is a specific technique to amplify human gene sequences from complex heterologous DNAs by using human Alu primers.
出处
《军事医学科学院院刊》
CSCD
北大核心
1998年第3期199-202,共4页
Bulletin of the Academy of Military Medical Sciences
基金
国家自然科学基金