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携带兔β-珠蛋白基因编码序列的SV40重组体。

SV40 recombinants carrying rabbit beta-globin gene coding sequences.

作者信息

Hamer D H, Smith K D, Boyer S H, Leder P

出版信息

Cell. 1979 Jul;17(3):725-35. doi: 10.1016/0092-8674(79)90279-4.

Abstract

We have constructed and propagated two SV40 recombinants in which different portions of the viral late gene region are replaced by a rabbit beta-globin coding sequence derived from a cloned cDNA (Maniatis et al., 1976). One of these recombinants, which retains the promoter, leader and intervening sequences for a viral late mRNA, directs the synthesis of a stable SV40-globin hybrid transcript. Using a sensitive radioimmunoassay, we have shown that this globin RNA is translated in infected monkey cells. A second recombinant, which retains the late region promoter but lacks the RNA splicing region, produces neither a stable globin transcript nor any detectable beta-globin. These experiments indicate that some sequence within a 500 bp SV40 segment, presumably the splicing region, is required for the accumulation of stable mRNA. They also demonstrate how hybrid transducing viruses can be used both to characterize viral regulatory sequences and to produce new gene products in cultured cells.

摘要

我们构建并繁殖了两种SV40重组体,其中病毒晚期基因区域的不同部分被源自克隆cDNA的兔β-珠蛋白编码序列所取代(Maniatis等人,1976年)。这些重组体中的一个保留了病毒晚期mRNA的启动子、前导序列和间隔序列,指导合成稳定的SV40-珠蛋白杂交转录本。使用灵敏的放射免疫测定法,我们已表明这种珠蛋白RNA在受感染的猴细胞中被翻译。第二个重组体保留了晚期区域启动子但缺乏RNA剪接区域,既不产生稳定的珠蛋白转录本,也不产生任何可检测到的β-珠蛋白。这些实验表明,500 bp SV40片段内的某些序列,大概是剪接区域,是稳定mRNA积累所必需的。它们还证明了杂交转导病毒如何可用于表征病毒调控序列以及在培养细胞中产生新的基因产物。

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