Zheng C, Baum B J, Iadarola M J, O'Connell B C
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, USA.
Nat Biotechnol. 2000 Feb;18(2):176-80. doi: 10.1038/72628.
A replication-deficient recombinant adenovirus encoding luciferase was constructed using 5' and 3' long terminal repeat (LTR) sequences of the Moloney murine leukemia virus. Gene expression was observed in cultured cells in vitro and in submandibular gland, cortex, and caudate nucleus for as long as three months in vivo. The vector integrated randomly into the genome of both dividing and nondividing cells as determined by fluorescence in situ hybridization (FISH) (10-15% of cells in vitro and 5% in rat spleen in vivo), gene walking, Southern hybridization, and polymerase chain reaction (PCR), in the absence of transcomplementing reverse transcriptase or integrase activity. The new vector combines the high titer and versatility of adenoviral vectors with the long-term gene expression and integration of retroviral vectors.
利用莫洛尼鼠白血病病毒的5'和3'长末端重复序列（LTR）构建了一种编码荧光素酶的复制缺陷型重组腺病毒。在体外培养细胞以及体内颌下腺、皮质和尾状核中观察到基因表达长达三个月。通过荧光原位杂交（FISH）（体外10 - 15%的细胞，体内大鼠脾脏中5%的细胞）、基因步移、Southern杂交和聚合酶链反应（PCR）确定，该载体随机整合到分裂细胞和非分裂细胞的基因组中，且不存在反式互补逆转录酶或整合酶活性。这种新型载体将腺病毒载体的高滴度和多功能性与逆转录病毒载体的长期基因表达和整合特性结合在一起。
