Gerenday A, Shih K M, Herman C C, Fallon A M
Department of Entomology, University of Minnesota, St. Paul, MN 55108, USA.
Arch Insect Biochem Physiol. 2001 Jan-Feb;46(1-2):19-25. doi: 10.1002/arch.4.
Hydroxyurea-resistant Aedes albopictus mosquito cells were selected by incremental exposure of unmutagenized cells to hydroxyurea concentrations ranging from 0.1 to 8 mM. Clonal populations that had become 40-fold more resistant to hydroxyurea than wild-type cells varied in morphology, and their growth rate decreased to a;45 h doubling time, relative to an 18 h doubling time in unselected cells. At this level of resistance, the cells remained diploid, with a modal chromosome number of 6. When labelled with (35)S[methionine/cysteine], clone HU1062, which grew in the presence of 8 mM hydroxyurea, overproduced a labeled protein with the approximate size of the 45,000 dalton M2 subunit of ribonucleotide reductase. Consistent with this observation, ribonucleotide reductase activity in HU-1062 cells was approximately 10-fold higher than in wild-type control cells. This is the first example of an hydroxyurea-resistant insect cell line. [Originally published in Volume 34, Archives of Insect Biochemistry and Physiology, 34:31-41 (1997).]
通过将未诱变的细胞逐步暴露于浓度范围为0.1至8 mM的羟基脲中来筛选对羟基脲具有抗性的白纹伊蚊细胞。对羟基脲的抗性比野生型细胞高40倍的克隆群体在形态上有所不同，并且它们的生长速率降低至45小时的倍增时间，而未筛选细胞的倍增时间为18小时。在这种抗性水平下，细胞保持二倍体状态，众数染色体数为6。当用(35)S[甲硫氨酸/半胱氨酸]标记时，在8 mM羟基脲存在下生长的克隆HU1062过量产生一种标记蛋白，其大小约为核糖核苷酸还原酶45,000道尔顿M2亚基。与该观察结果一致，HU-1062细胞中的核糖核苷酸还原酶活性比野生型对照细胞高约10倍。这是抗羟基脲昆虫细胞系的首个实例。[最初发表于《昆虫生物化学与生理学档案》第34卷，34:31 - 41(1997)]
