For gene transfer in fish, the cellular or viral promoters derived from mammalian are probably the only choice to use for the time being. This is due to the fact that the promoter of homologous transcriptional regulat...For gene transfer in fish, the cellular or viral promoters derived from mammalian are probably the only choice to use for the time being. This is due to the fact that the promoter of homologous transcriptional regulatory sequences is absent in fish.In this study, the bacterial gene for chloramphenicol acetyltransferase (CAT) under the control of the early promoter from simian virus 40 (SV40)was used to transfect the cultured kidney cells of grass carp and efficient expression was obtained. It is suggested that the mammalian viral SV40 promoter can be used for the expression of the foreign gene in transgenic fish.展开更多
萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断...萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断中一段长为 4 9bp的 DNA序列 ,且荧光素酶基因 c DNA编码区的 5’端含有一个原核细胞基因表达调控区相似序列 .将 p GL2 中的荧光素酶基因 c DNA(L uc)克隆入真核表达载体 p ED中 ,构建成 p ED- Luc.该载体经表达测定后 ,只在真核细胞中表达 ,在原核细胞中不表达 .这说明荧光素酶基因 c DNA编码区的 5’端的原核细胞基因表达调控区相似序列不具表达有活性的荧光素酶的功能 .经 DNA序列对比研究 ,认为 p GL2 含有的 SV4 0基因组 Hind B片段 5 12 3bp～ 5 171bp长 4 9bp的 DNA序列可能就是 Hind B片段的原核增强子功能的决定序列 .展开更多
文摘For gene transfer in fish, the cellular or viral promoters derived from mammalian are probably the only choice to use for the time being. This is due to the fact that the promoter of homologous transcriptional regulatory sequences is absent in fish.In this study, the bacterial gene for chloramphenicol acetyltransferase (CAT) under the control of the early promoter from simian virus 40 (SV40)was used to transfect the cultured kidney cells of grass carp and efficient expression was obtained. It is suggested that the mammalian viral SV40 promoter can be used for the expression of the foreign gene in transgenic fish.
文摘萤火虫荧光素酶基因 c DNA真核表达载体 p GL2 可在原核细胞中高效表达 .该载体中含有 SV4 0早期启动子及荧光素酶基因 c DNA 5’端总长共 75 8bp的 DNA碱基序列 ,经计算机分析发现 ,在 SV4 0早期启动子中含有 SV4 0基因组 Hind B片断中一段长为 4 9bp的 DNA序列 ,且荧光素酶基因 c DNA编码区的 5’端含有一个原核细胞基因表达调控区相似序列 .将 p GL2 中的荧光素酶基因 c DNA(L uc)克隆入真核表达载体 p ED中 ,构建成 p ED- Luc.该载体经表达测定后 ,只在真核细胞中表达 ,在原核细胞中不表达 .这说明荧光素酶基因 c DNA编码区的 5’端的原核细胞基因表达调控区相似序列不具表达有活性的荧光素酶的功能 .经 DNA序列对比研究 ,认为 p GL2 含有的 SV4 0基因组 Hind B片段 5 12 3bp～ 5 171bp长 4 9bp的 DNA序列可能就是 Hind B片段的原核增强子功能的决定序列 .
