チミジンキナーゼの新しいタイプの変異とアシクロビル耐性単純ヘルペスウイルス

胸苷激酶的新型突变与耐阿昔洛韦单纯疱疹病毒

• 批准号: 10770378
• 负责人: 西條 政幸
• 金额: $ 1.22万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10770378/
• 关键词: 単純ヘルペスウイルス; チミジン燐酸化酵素; アシクロビル; 耐性; チミジンリン酸化酵素; アシクロピール

1)アシクロビル(ACV)感受性単純ヘルペスウイルス(HSV-1 TAS)からのACV耐性HSV-1の作製:ACV濃度を徐々に高めた培地でHSV-1 TASを培養し,ACV耐性HSV-1を24株作製した.2)ACV耐性HSV-1のチミジンリン酸化酵素(TK)遺伝子の塩基配列:24株のACV耐性HSV-1のうち,8株の遺伝子は430-436番目のGが7塩基続く部位にGが1塩基挿入され,Gが8塩基続く変異を起こしていた.2株は548-553番目のCが6連続する塩基配列のうち,1塩基のCが欠損する変異を起こしていた.また,2株は1071-1074番目に位置する4連続したCから1塩基のCが欠損する変異を起こしていた.このように24株のうち,12株はGまたはCが連続する塩基配列に,塩基の挿入または欠損という変異を伴っていた.その他の12株においては,8株がさまざまな部位に1アミノ酸変異をもつTK蛋白を,3株が1塩基置換による早期ストップコドン出現による短いTK蛋白を,1株が連続する塩基配列の欠損による短いTK蛋白を発現した.3)TK活性とACVリン酸化酵素活性の測定:24株のACV耐性HSV-1のうち,2株がTK活性を親株とほぼ同等に発現するTK変異株で,残りの22株はTK欠損株であった.2株のTK変異株はともに1アミノ酸変異をもつTK蛋白を発現していた.4)結論:ACV耐性HSV-1のほぼ半分は,TK遺伝子における連続するGあるいはCの部位に1塩基の欠損・挿入によるACV耐性を獲得し,30%はTK蛋白のアミノ酸変異によるACV耐性を獲得する.また,1アミノ酸変異によるACV耐性株8株のうち,4株はATP-binding siteやnucleotide-binding site以外の部位のアミノ酸変異を有していた.つまり,それら以外の部位におけるアミノ酸変異もTK活性やACVリン酸化活性に影響をあたえる.

1)(ACV) Susceptibility to Pure Species (HSV-1 TAS) ACV tolerance HSV-1 HSV-1 TAS was cultured at high ACV concentrations, and 24 ACV resistant HSV-1 strains were produced. 2)ACV resistant HSV-1 and TK gene mapping: 24 ACV resistant HSV-1 strains were isolated, 8 ACV resistant HSV-1 strains were isolated from 430-436 HSV-1 strains, 8 ACV resistant HSV-1 strains were isolated from 7 HSV-1 strains, 8 ACV resistant HSV-1 strains were isolated from 7 HSV-1 strains, G: 8 base: 6 base: 1 base 2 plants 1071-1074 positions 4 consecutive years C 1 year C 1 year C 2 year C 3 year C 4 year C 3 year C 3 year C 4 year C 4 year C 3 year C 4 year C 4 year C 3 year C 4 year C 6 year C year C year 6 year C year 6 year 6 year 24 of these plants were found to be deficient in G, C and D, and 12 were found to be deficient in G, C and D. Among the 12 strains, 8 strains showed 1 loss of TK protein at the locus of HSV-1, 3 strains showed 1 loss of TK protein at the locus of HSV-1, 3 strains showed 1 loss of TK protein at the locus of HSV-1, 1 strain showed 1 loss of TK protein at the locus of HSV-1, 3 strains showed 1 loss of TK protein at the locus of HSV-1, 2 strains showed 2 loss of TK protein at the locus of HSV-1, 2 strains showed 2 loss of TK protein at the locus of HSV-1, 3 strains showed 2 loss of TK protein at the locus of HSV-1, 2 strains showed 2 loss of TK protein at the locus of HSV-1, 3 strains showed 2 loss of TK protein at the locus of HSV-1, 4 strains showed 2 loss of TK protein at the locus of HSV-1. Conclusion: ACV tolerance was achieved in HSV-1,TK protein in C and TK protein in HSV-1, and 30% of TK protein in HSV-1 and TK protein in C. 1 ACV resistant strain was resistant to acid variation, 8 ACV resistant strains were resistant to acid variation, 4 ACV resistant strains were resistant to acid variation except ATP-binding site nucleotide-binding site. The activity of TK and ACV in different parts of the body is affected by acid.

项目成果

Masayuki Saijo et al.: "Nucleotide sequence of thymidime kinase gene of sequential acyclovir-resistant herpes simplex virus type 1 recovered from a child with WAS."Journal of Medical Virology. 58. 387-393 (1999)

Masayuki Saijo 等人：“从患有 WAS 的儿童身上发现的连续阿昔洛韦耐药的 1 型单纯疱疹病毒的胸苷激酶基因的核苷酸序列。”医学病毒学杂志。