Law Lane K, Davidson Beverly L
Program in Gene Therapy, Program in Genetics, Department of Internal Medicine, Neurology, and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
J Virol. 2002 Jan;76(2):656-61. doi: 10.1128/jvi.76.2.656-661.2002.
Prior work by members of our laboratory and others demonstrated that adenovirus serotype 30 (Ad30), a group D adenovirus, exhibited novel transduction characteristics compared to those of serotype 5 (Ad5, belonging to group C). While some serotype D adenoviruses bind to the coxsackie-adenovirus receptor (CAR), the ability of Ad30 fiber to bind CAR is unknown. We amplified and purified Ad30 and cloned the Ad30 fiber by overlap PCR. Alignment of Ad30 fiber with Ad3, Ad35, Ad5, Ad9, and Ad17 revealed that Ad30, like Ad9 and Ad17, has a shortened fiber sequence relative to that of Ad5. The knob region of fiber was 45% identical to that of the Ad5 knob regions. We made a chimeric recombinant virus (Ad5GFPf30) in which the Ad5 fiber (amino acids [aa]47 to 582) was replaced with Ad30 fiber sequences (aa 46 to 372), and CAR-mediated viral entry was determined on CAR-expressing Chinese hamster ovary (CHO) cells. While CAR expression significantly increased Ad5GFP-mediated transduction in CHO cells (from 1 to 36%), it did not enhance Ad5GFPf30 gene transfer. Binding of radiolabeled Ad5GFPf30 or Ad30 wild-type virus was also not improved by the expression of CAR. These results suggest that Ad30 fiber is distinct from Ad5, Ad9, and Ad17 fibers in its inability to direct transduction via CAR.
我们实验室成员及其他人员之前的研究表明,D组腺病毒血清型30(Ad30)与C组腺病毒血清型5(Ad5)相比,具有独特的转导特性。虽然一些D组腺病毒血清型可与柯萨奇病毒-腺病毒受体(CAR)结合,但Ad30纤维与CAR结合的能力尚不清楚。我们扩增并纯化了Ad30,并通过重叠PCR克隆了Ad30纤维。将Ad30纤维与Ad3、Ad35、Ad5、Ad9和Ad17进行比对,发现Ad30与Ad9和Ad17一样,相对于Ad5,其纤维序列缩短。纤维的球状结构域与Ad5球状结构域的同源性为45%。我们构建了一种嵌合重组病毒(Ad5GFPf30),其中Ad5纤维(氨基酸[aa]47至582)被Ad30纤维序列(aa 46至372)取代,并在表达CAR的中国仓鼠卵巢(CHO)细胞上测定了CAR介导的病毒进入情况。虽然CAR表达显著增加了Ad5GFP介导的CHO细胞转导(从1%增至36%),但并未增强Ad5GFPf30的基因转移。CAR的表达也未改善放射性标记的Ad5GFPf30或Ad30野生型病毒的结合。这些结果表明,Ad30纤维在通过CAR介导转导方面与Ad5、Ad9和Ad17纤维不同。