生物化学与生物物理进展
生物化學與生物物理進展
생물화학여생물물리진전
PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2009年
3期
358-363
,共6页
杨鹏辉%安文琪%石辛甫%段跃强%罗德炎%张鹏飞%唐冲%邢丽%张玉静%刘秀梵%王希良
楊鵬輝%安文琪%石辛甫%段躍彊%囉德炎%張鵬飛%唐遲%邢麗%張玉靜%劉秀梵%王希良
양붕휘%안문기%석신보%단약강%라덕염%장붕비%당충%형려%장옥정%류수범%왕희량
B型流感病毒%反向遗传学%8质粒拯救系统%重组流感病毒
B型流感病毒%反嚮遺傳學%8質粒拯救繫統%重組流感病毒
B형류감병독%반향유전학%8질립증구계통%중조류감병독
influenza B virus%reverse genetics%eight-plasmid system%reassorted influenza virus
以冷适应、温度敏感、减毒的B/Ann Arbor/1/66流感病毒株作为重配病毒骨架,对其6个内部基因片段进行了全基因合成,同时人工引入9个氨基酸突变.构建了8个基因的拯救载体,经测序获得序列准确的拯救质粒,命名为:pAB121-PB1,pAB122-PB2,pAB123-PA,pAB124-HA,pAB125-NP,pAB126-NA,pAB127-M和pAB128-NS.在成功拯救冷适应A型流感病毒的基础上,利用反向遗传学技术成功获救了具有感染性的重配B型流感病毒株,命名为rMDV-B.该重配病毒株以B/Ann Arbor/1/66为病毒骨架,其中HA和NA来源于2006～2007年当年流行株B/Malaysia/2506/2004.rMDV-B在鸡胚尿囊液和MDCK细胞中的HA效价可达1:64～1:512.实验结果暗示:从单一供体病毒株可以产生有效的减毒活B型流感病毒疫苗候选株,能够为将来人用流感疫苗的设计提供可借鉴的模型.
以冷適應、溫度敏感、減毒的B/Ann Arbor/1/66流感病毒株作為重配病毒骨架,對其6箇內部基因片段進行瞭全基因閤成,同時人工引入9箇氨基痠突變.構建瞭8箇基因的拯救載體,經測序穫得序列準確的拯救質粒,命名為:pAB121-PB1,pAB122-PB2,pAB123-PA,pAB124-HA,pAB125-NP,pAB126-NA,pAB127-M和pAB128-NS.在成功拯救冷適應A型流感病毒的基礎上,利用反嚮遺傳學技術成功穫救瞭具有感染性的重配B型流感病毒株,命名為rMDV-B.該重配病毒株以B/Ann Arbor/1/66為病毒骨架,其中HA和NA來源于2006～2007年噹年流行株B/Malaysia/2506/2004.rMDV-B在鷄胚尿囊液和MDCK細胞中的HA效價可達1:64～1:512.實驗結果暗示:從單一供體病毒株可以產生有效的減毒活B型流感病毒疫苗候選株,能夠為將來人用流感疫苗的設計提供可藉鑒的模型.
이랭괄응、온도민감、감독적B/Ann Arbor/1/66류감병독주작위중배병독골가,대기6개내부기인편단진행료전기인합성,동시인공인입9개안기산돌변.구건료8개기인적증구재체,경측서획득서렬준학적증구질립,명명위:pAB121-PB1,pAB122-PB2,pAB123-PA,pAB124-HA,pAB125-NP,pAB126-NA,pAB127-M화pAB128-NS.재성공증구랭괄응A형류감병독적기출상,이용반향유전학기술성공획구료구유감염성적중배B형류감병독주,명명위rMDV-B.해중배병독주이B/Ann Arbor/1/66위병독골가,기중HA화NA래원우2006～2007년당년류행주B/Malaysia/2506/2004.rMDV-B재계배뇨낭액화MDCK세포중적HA효개가체1:64～1:512.실험결과암시:종단일공체병독주가이산생유효적감독활B형류감병독역묘후선주,능구위장래인용류감역묘적설계제공가차감적모형.
A cold-adapted (ca), temperature sensitive (ts), live attenuated influenza B virus strain B/Ann Arbor/1/66 was chosen for influenza virus rescue research, in which six internal gene segments, PB1, PB2, PA, NP, M, NS, were fully synthesized and nine amino acid substitutions were artificially alter by human intervention. The resultant B/Ann Arbor/1/66 plasmids were named as pAB121-PB1, pAB122-PB2, pAB123-PA, pAB124-HA, pAB125-NP, pAB126-NA, pAB127-M and pAB128-NS, respectively. A recombinant influenza A virus was previously generated entirely from cloned cDNA. An infectious recombinant influenza B virus was generated here, and designated as rMDV-B, by plasmid-based reverse genetics. The rMDV-B virus contained HA and NA genes from an epidemic influenza B vires strain B/Malaysia/2506/2004 in the background of internal genes derived from influenza B virus strain B/Ann Arbor/1/66. HA titer of rMDV-B in MDCK cells and embryonated chicken eggs ranged from 1 : 64 to 1 : 512. The results may allow an effective live influenza B vaccine to be produced from a single master strain, providing a model for the design of future live human influenza vaccines.
