Bradford C S, Miller A E, Toumadje A, Nishiyama K, Shirahata S, Barnes D W
Department of Biochemistry and Biophysics, Environmental Health Sciences Center, Oregon State University, Corvallis 97331-7305, USA.
Mol Mar Biol Biotechnol. 1997 Dec;6(4):279-88.
The Japanese pufferfish (genus Fugu), which possesses a highly compact genome, is becoming a popular model among those interested in sequencing and mapping the genomes of higher vertebrates. Although genomic libraries have been derived and used to study the molecular biology of Fugu, biological material derived from the living organism is difficult to obtain for laboratories distant from the Asian Pacific. We have established cell cultures from two Fugu species: kusafugu, Fugu niphobles, and torafugu, F. rubripes. Cultures derived from F. niphobles fry and F. rubripes eye have been passaged more than 60 times over the course of one year, representing approximately 180 population doublings. Proliferating cultures were also initiated from F. rubripes brain, liver, fin, spleen, kidney, swimbladder, and muscle. Karyotype analyses indicated that F. rubripes eye-derived cells possessed a chromosome number in the diploid range; F. niphobles fry cells were slightly hyperploid. Flow cytometry confirmed that the relative amounts of DNA present in cultured cells from both Fugu species were similar to that measured in blood cells collected from F. rubripes, and approximately one-seventh of that measured in diploid human cells. Telomerase activity was easily detectable in lysates prepared from F. niphobles fry cells and F. rubripes eye cells, consistent with the notion that these cultures are capable of indefinite proliferation.
日本河豚（鲀属）拥有高度紧凑的基因组，正成为那些对高等脊椎动物基因组测序和图谱绘制感兴趣的人所青睐的模式生物。尽管已经构建了基因组文库并用于研究河豚的分子生物学，但对于远离亚太地区的实验室来说，从活体生物获取生物材料却很困难。我们已经从两种河豚中建立了细胞培养物：虎河豚，黑斑叉鼻鲀，和虎河豚，红鳍东方鲀。来自黑斑叉鼻鲀幼鱼和红鳍东方鲀眼睛的培养物在一年时间里传代超过60次，大约相当于180次群体倍增。还从红鳍东方鲀的脑、肝、鳍、脾、肾、鳔和肌肉中启动了增殖培养。核型分析表明，红鳍东方鲀眼睛来源的细胞染色体数目处于二倍体范围内；黑斑叉鼻鲀幼鱼细胞略为超倍体。流式细胞术证实，两种河豚培养细胞中DNA的相对含量与从红鳍东方鲀采集的血细胞中测得的相似，约为二倍体人类细胞中测得含量的七分之一。在黑斑叉鼻鲀幼鱼细胞和红鳍东方鲀眼睛细胞制备的裂解物中很容易检测到端粒酶活性，这与这些培养物能够无限增殖的观点一致。
