CAJ | 학술논문

为了构建RNA病毒样颗粒（virus-like particles ，VLP）表达系统pET28a（＋）-MS，将插入的外源基因诱导表达获得相应的RNA标准物质病毒样颗粒，将MS2噬菌体包膜蛋白和成熟蛋白基因插入表达质粒pET-28a（＋），构建pET28a（＋）-MS重组子，将带有包装位点序列的外源 RNA对应的 cDNA插入成熟蛋白基因的下游，经过诱导表达含外源 RNA病毒样颗粒。结果表明，pET28a（＋）-MS能表达活性的包膜蛋白（49．8 ku）和成熟蛋白（14．4 ku），能将外源基因 RNA包装成病毒样颗粒，该 VLP长度约1500 bp， RT-PCR反应性优良，能耐受RNase的降解，在血清中37℃经1个月未有明显降解。
위료구건RNA병독양과립（virus-like particles ，VLP）표체계통pET28a（＋）-MS，장삽입적외원기인유도표체획득상응적RNA표준물질병독양과립，장MS2서균체포막단백화성숙단백기인삽입표체질립pET-28a（＋），구건pET28a（＋）-MS중조자，장대유포장위점서렬적외원 RNA대응적 cDNA삽입성숙단백기인적하유，경과유도표체함외원 RNA병독양과립。결과표명，pET28a（＋）-MS능표체활성적포막단백（49．8 ku）화성숙단백（14．4 ku），능장외원기인 RNA포장성병독양과립，해 VLP장도약1500 bp， RT-PCR반응성우량，능내수RNase적강해，재혈청중37℃경1개월미유명현강해。
To construct virus-like particles(VLP)expression platform pET28a(+)-MS,the inserted foreign gene is induced to express the VLP containing corresponding RNA standard material.The envelope and mature protein genes of MS2 bacteriophages were inserted into plasmid pET-28a(+).The cDNA corre-sponding an exogenous RNA with package sequence was inserted into mature protein gene downstream, and the recombinant pET28a(+)-MS was induced to express exogenous RNA containing VLP.The results showed that,pET28a(+)-MS can express active envelope protein (49.8 ku)and mature protein (14.4 ku),the foreign RNA was packaged into VLP.the VLP is about 1 500 bp long,RT-PCR has excellent re-activity,can withstand the degradation by RNase in serum 3 7 ℃ 1 month and no significant degradation de-tected.