USE OF RETROVIRAL SHUTTLE VECTOR FOR INFECTION OF ONCOGENES INTO HUMAN CELLS

病毒携带正义或反义方向的 已经产生了以下基因：v-raf、c-raf、v-Ha-ras、c-raf。 myc（小鼠）和PDGF A链（正常和羧基缺失的 顺反子）。 ras、v-raf和PDGF A链的正义构建体 发现在小鼠3 T3细胞上产生转化灶。 最 在具有相对短的乳腺的裸鼠中产生肿瘤。 用Zip-Ha-ras病毒感染永生化人支气管 上皮细胞（Beas 12）。 皮下注射后5周内 将500万个细胞感染到裸鼠体内， 80%的老鼠 这些培养细胞的核型显示 他们是人类的起源。 用ras反义Zip-Ha-sar病毒感染TBE-1细胞 （转化的原代人支气管上皮细胞 因为它们是v-Ha-ras癌基因所必需的）。 我们的结论 从这个实验可以看出Ha-ras基因的功能是必需的 维持TBE-1细胞增殖，因为感染的细胞， 应该在G418的存在下生长，而不是因为 功能的反义废除。 Zip-Ha-sar感染的细胞 缺乏先前引入的v-Ha-ras mRNA， 预期的5.4-kb mRNA代表SAR转录物。

Viruses carrying either sense or anti-sense orientations of the following genes have been produced: v-raf, c-raf, v-Ha-ras, c- myc(mouse), and PDGF A chain (normal and carboxyl-deleted cistrons). The sense constructs of ras, v-raf, and PDGF A chain were found to produce transformed foci on mouse 3T3 cells. Most produced tumors in nude mice with relatively short latencies. Zip-Ha-ras virus was used to infect immortalized human bronchial epithelial cells (Beas 12). Within 5 weeks from subcutaneous infection of 5 million cells into nude mice, tumors appeared in 80% of the mice. The karyotype of these cultured cells shows them to be human in origin. Zip-Ha-sar virus (ras anti-sense) were used to infect TBE-1 cells (primary human bronchial epithelial cells which were transformed after they were essential with a v-Ha-ras oncogene). We conclude from this experiment that the Ha-ras gene function is necessary to maintain TBE-1 cell proliferation since infected cells, which should grow in the presence of G418, do not as a consequence of anti-sense abrogation of function. The Zip-Ha-sar infected cells lack the previously introduced v-Ha-ras mRNA and have instead the expected 5.4-kb mRNA representing the sar transcript.