微粒沈積均勻度的量測與應用

Abstract

常見基因轉殖的基因載入模式，包括︰基因槍、直接肌肉注射、複合基因、電穿孔、細菌或病毒感染等。其中以基因槍的基因載入模式，對細胞危害性低、所需的載入物質劑量低、可載入的DNA劑量大、以及可廣泛的應用在各種體內和體外的培養細胞等優點，使得基因槍成為生物技術上的新趨勢。 本研究針對影響基因槍之金微粒沈積均勻度與收集效率的因子，像是氦氣出口大小( 0.05, 0.1, 0.5, 5, 20 mm)、出口位置( 0, 15, 25 mm)、氦氣氣流壓力(100, 150, 250, 400, 600 psi)、錐狀體長度(5, 15, 35, 65, 105, 155 mm)、金微粒沉積面積的直徑(10, 15, 25, 30, 50 mm)、筒狀體長度(25, 35, 45, 65, 95, 155 mm)等，以探討對基因轉殖效率的影響，進而找出最佳化的轉殖效度：氦氣出口大小0.05 mm、出口位置0 mm、氦氣氣流壓力600 psi、錐狀體長度65 mm、金微粒沉積面積的直徑50 mm。改良後的基因槍收集效率增加為原先設計的5.2倍、均勻度提升了2.5倍、轉殖效度提升了17.3倍。

The ways of gene deliver during gene transfection included gene gun, injection directly into the muscle, complex DNAs, electroporation, bacterial systems, and viral delivery systems. Among them, the advantages of the gene gun was below: low harmfulness for cell, only small amounts of DNA for requirement, but large DNA fragments may be transferred, applicable to both in vitro and in vivo transformation. So the gene gun will become a new trend in biological technology. In this research, we confer relationship between uniformity and slot width (0.05, 0.1, 0.5, 5, 20 mm), slot height (0, 15, 25 mm), helium pressure (100, 150, 250, 400, 600 psi), transition cone (5, 15, 35, 65, 105, 155 mm), diameter of target filter (10, 15, 25, 30, 50 mm), column height (25, 35, 45, 65, 95, 155 mm). The best transfection occurs when slot width is 0.05 mm, slot height is 0 mm, transition cone is 65 mm and diameter of target filter is 50 mm. Collection efficiency is 5.2 times than original design, uniformity is 2.5 times than original design and transfection is 17.3 times than original design.