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Summary The embryonic cell line, GV1, from Manduca sexta was transiently transfected with DNA constructs of the Drosophila hsp70 promoter fused to either a β-galactosidase (pXH70ZT) or a chloramphenicol acetyl transferase (HSP-CAT-1) reporter gene
using lipofectin. Optimal cell density, DNA:lipofectin ratio, and time of incubation were varied to determine the optimal
conditions: 2 × 105 cells/ml, 1:3, and 5 h. Under these conditions, the transfection efficiency was about 40%.
Heat inducibility of two hsp70 constructs was compared. The HSP-CAT-1, containing 1127 bp of upstream sequence, was more sensitive
to heat shock than that of pXH70ZT, containing only 194 bp of upstream sequence. Thus, the 1127 bp hsp70 promoter appears
to be a better inducible promoter in these cells. A 2 kb fragment of the proximal promoter region of the MHR3 gene containing
a putative ecdysone response element was shown to be responsive to 20-hydroxyecdysone after its transfection into these cells. 相似文献
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The efficiency of dendrosomes (novel dendritic spheroidal nanoparticle gene porters) were assessed in transferring wild-type
p53 cDNA into two human acute lymphoblastic leukemia cell lines (MOLT-4 and CCRF-CEM) derived from T cells and erythroleukemic
cell line K562. Flow cytometric studies showed a 65% and 45% enhancement in apoptosis and necrosis of K562 and CCRF-CEM cells
transfected with complex of dendrosomes and wild-type p53 cDNA in comparison to controls. The cytotoxicity studies on T lymphoma
cells revealed that dendrosomes have a low cytotoxicity in comparison to lipofectin. 相似文献
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特异性扩增家鸡卵清蛋白基因上游调控序列 1340bp~ +16 5 5bp片段和第一内含子 +49bp~ +16 5 5bp片段 ,去除pG FP N2载体自身的CMV启动子 ,分别构建了P2.9koval GFP和P1.5koval GFP两种表达载体 ,经测序和酶切鉴定表达载体构建正确。采用脂质体转染法分别将这两种载体、pGFP N2 (阳性对照 )质粒及阴性对照转染鸡原代输卵管上皮细胞和中国仓鼠卵巢细胞。用荧光倒置显微镜观测绿色荧光蛋白的表达。结果表明 :两种表达质粒在鸡原代输卵管上皮细胞和中国仓鼠卵巢细胞中都可以表达荧光蛋白。结果既显示卵清蛋白第一内含子对基因的表达起到一定的调控作用 ,也显示卵清蛋白启动子对输卵管上皮细胞和卵巢细胞不存在特异性 ,并且不存在种属差异性。 相似文献
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