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将基因转入细胞常用逆病毒和AAV及脂质体作为载体,但这些载体对处于非分裂状态细胞如人骨髓是无效的。SV40因其高传导效率和广泛的宿主范围,近几年来成为人们注目的载体,它能将基因有效地转入人体各种细胞包括骨髓细胞。SV40为环状双DNA,属乳多空病毒,大小为5.2kb。最初在美国作为脊髓灰质炎疫苗污染物成分被发现,但随机检查发现对人类并不致病。本文证实了SV40转导3.8kb MDR基因的可行性,并发现了经体外包装后的SV40假病毒粒子—pSM1。 选用COS做包装细胞,其他细胞有CMT4,K562,NIH-3T3,P388,MEL。用RT-PCR,罗丹明-123拒染法和流式细胞分析法检测细胞基因的转录
Transfection of genes into cells commonly uses retroviruses and AAV and liposomes as vectors, but these vectors are ineffective for cells in non-dividing state such as human bone marrow. Due to its high conduction efficiency and wide host range, SV40 has become an attractive carrier in recent years. It can transfer genes efficiently into various human cells, including bone marrow cells. SV40 is a circular double DNA, is a papvirus, the size of 5.2kb. Originally found in the United States as a polio ingredient in polio vaccines, randomized studies have found no risk to humans. This article confirms the feasibility of SV40 transduction of the 3.8 kb MDR gene and the discovery of the in vitro-packaged SV40 pseudovirions-pSM1. COS used to make packaging cells, other cells are CMT4, K562, NIH-3T3, P388, MEL. RT-PCR, rhodamine-123 dyed assay and flow cytometry assay of cellular gene transcription