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刊物信息

期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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改构溶瘤腺病毒DNA序列分析研究

Study on DNA sequence analysis of a modified oncolytic adenovirus

分类号:R917
出版年·卷·期(页码):2020,40 (1):43-47
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的: 以改构的溶瘤腺病毒制品为研究对象,探讨DNA测序技术在基因治疗制品质量控制和质量标准制定中的作用。方法: 提取并纯化一种改构溶瘤腺病毒制品的基因组DNA,对其基因组关键区段、非关键区段进行测序分析,发现变异体后应用第一代和第二代DNA测序技术进行进一步分析和检测。结果: 关键区段DNA序列与理论序列一致;非关键区段DNA序列与理论序列相似度大于99.9%;发现1个高变异区段,进一步分析结果显示:缺失1个胞嘧啶(C)的变异体占比5/9(克隆/克隆);缺失2个胞嘧啶(C)的变异体占比1/9(克隆/克隆);插入1个胞嘧啶(C)的变异体占比1/9(克隆/克隆)。结论: 第一代和第二代DNA测序技术虽然各有优缺点,但只要相互组合使用得当,仍然可较好地用于基因治疗制品基因组的分析和质量控制。

-----英文摘要:---------------------------------------------------------------------------------------

Objective: To explore the application of DNA sequencing technology in quality control and quality standard establishment of gene therapy products by a case study of a modified oncolytic adenovirus product. Methods: The genomic DNA of a modified oncolytic adenovirus product was extracted and purified,and the key segments and non-key segments of the genome were sequenced and analyzed. Once mutant DNA segments were found,further detection and analysis were conducted by a combination of the first and second generation DNA sequencing technology. Results: The measured sequence of key DNA segments was consistent with the expected sequence. The similarity between the measured and expected DNA sequence for non-key segments was more than 99.9%. A variable region was found,and further analysis showed that the proportion of variant with a deletion of one cytosine(C) was 5/9(clones/clones),the proportion of variant with a deletion of two cytosine(C) was 1/9(clone/clone),the proportion of variant with an insert of one cytosine(C) was 1/9(clone/clone). Conclusion: Although the first and second generation of DNA sequencing technology has their own advantages and disadvantages,they can be well used in genomic analysis and quality control of gene therapy products by appropriate combination.

-----参考文献:---------------------------------------------------------------------------------------

[1] 徐疏梅.新一代DNA测序技术的应用与研究进展[J].徐州工程学院学报(自然科学版),2018,34(12):60 XU SM.Application and research progress of a new generation of DNA sequencing technology[J].J Xuzhou Inst Technol(Nat Sci Ed),2018,34(12):60
[2] ISMAIL AM,LEE JS.Adenoviromics:mining the human adenovirus species D genome[J].Front Microbiol,2018,11(9):2178
[3] SAHA B,PARKS RJ.Human adenovirus type 5 vectors deleted of early region 1(E1) undergo limited expression of early replicative E2 proteins and DNA replication in non-permissive cells[J].PLoS One,2017,12(7):e0181012
[4] SAVELYEVA I,DOBBELSTEIN M.Infection with E1B-mutant adenovirus stabilizes p53 but blocks p53 acetylation and activity through E1A[J].Oncogene,2011,30(7):865
[5] AMALFITANO A,HAUSER MA,HU H, et al.Production and characterization of improved adenovirus vectors with the E1,E2b,and E3 genes deleted[J].J Virol,1998,72(2):926
[6] HAUT LH,GILL AL,KURUPATI RK,et al.A partial E3 deletion in replication-defective adenoviral vectors allows for stable expression of potentially toxic transgene products[J].Hum Gene Ther Methods,2016,27(5):187
[7] MEI YF,WU H,HULTENBY K,et al.Complete replication-competent adenovirus 11p vectors with E1 or E3 insertions show improved heat stability[J].Virology,2016,497:198
[8] WOLD WS,CLADARAS C,DEUTSCHER SL,et al.The 19-kDa glycoprotein coded by region E3 of adenovirus.Purification,characterization,and structural analysis[J].J Biol Chem,1985,260(4):2424
[9] FRIEDMAN JM,HORWITZ MS.Inhibition of tumor necrosis factor alpha-induced NF-kappa B activation by the adenovirus E3-10.4/14.5K complex[J].J Virol,2002,76(11):5515
[10] SORIANO AM,CRISOSTOMO L,MENDEZ M,et al.Adenovirus 5 E1A interacts with E4orf3 to regulate viral chromatin organization[J].J Virol,2019,93(10).pii:e00157
[11] CAI Z,LV H,CAO W,et al.Targeting strategies of adenovirus-mediated gene therapy and virotherapy for prostate cancer(Review)[J].Mol Med Rep,2017,16(5):6443
[12] 韩春梅,刘兰,杨靖清,等.重组人粒细胞刺激因子的N端氨基酸序列异质性分析[J].药物分析杂志,2018,38(11):1887 HAN CM,LIU L,YANG JQ,et al.Analysis of N-terminal amino acid sequence heterogeneity of recombinant human granulocyte colony-stimulating factor[J].Chin J Pharm Anal,2018,38(11):1887
[13] JIN X,JIANG Q,CHEN Y,et al.Similarity/dissimilarity calculation methods of DNA sequences:a survey[J].J Mol Graph Model,2017,76:342
[14] VOLLGER MR,LOGSDON GA,AUDANO PA,et al.Improved assembly and variant detection of a haploid human genome using single-molecule,high-fidelity long reads[J].Ann Hum Genet,2019.doi:10.1111/ahg.12364
[15] 李艳慧,张少强.DNA测序技术及其拼接算法综述[J].天津师范大学学报(自然科学版),2018,38(5).doi:10.19638/j.issn1671-1114.20180501 LI Yanhui,ZHANG Shaoqiang.Overview of DNA sequencing techniques and corresponding assembly algorithms[J].J Tianjin Norm Univ(Nat Sci Ed),2018,38(5).doi:10.19638/j.issn1671-1114.20180501

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