Girones R, Cote P J, Hornbuckle W E, Tennant B C, Gerin J L, Purcell R H, Miller R H
Hepatitis Viruses Section, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1989 Mar;86(6):1846-9. doi: 10.1073/pnas.86.6.1846.
Woodchuck hepatitis virus (WHV) DNA was cloned from viral particles obtained from the serum of a woodchuck with a naturally acquired infection. The complete nucleotide sequence of the virus genome was determined and found to be 3323 base pairs long. Transfection experiments demonstrated that the recombinant WHV DNA was infectious in each of 18 woodchucks tested and established a chronic carrier state in 1 of 13 neonates and 3 of 5 adult animals. WHV DNA from serum particles from the chronically infected neonate was cloned and the nucleotide sequence of three independent recombinants was compared directly with that of the input recombinant DNA. The consensus sequence of the three progeny genomes was identical to that of the parental DNA sequence. Therefore, transfection of woodchuck livers with recombinant WHV DNA induces active virus replication and gene expression and yields progeny genomes that are faithful copies of the input virus genome.
土拨鼠肝炎病毒(WHV)DNA是从一只自然感染的土拨鼠血清中获得的病毒颗粒中克隆出来的。测定了该病毒基因组的完整核苷酸序列,发现其长度为3323个碱基对。转染实验表明,重组WHV DNA在18只受试土拨鼠中的每一只中都具有感染性,并在13只新生动物中的1只和5只成年动物中的3只中建立了慢性携带状态。克隆了来自慢性感染新生动物血清颗粒的WHV DNA,并将三个独立重组体的核苷酸序列与输入的重组DNA的核苷酸序列直接进行比较。三个子代基因组的共有序列与亲本DNA序列相同。因此,用重组WHV DNA转染土拨鼠肝脏可诱导活跃的病毒复制和基因表达,并产生与输入病毒基因组忠实拷贝的子代基因组。