蚕业科学
蠶業科學
잠업과학
ACTA SERICOLOGICA SINICA
2010年
2期
350-353
,共4页
家蚕核型多角体病毒%orf126基因%C6基序%几丁质结合
傢蠶覈型多角體病毒%orf126基因%C6基序%幾丁質結閤
가잠핵형다각체병독%orf126기인%C6기서%궤정질결합
Bombyx moil nucleopolyhedrovirus%orf126 gene%C6 motif%Chitin binding
为了研究家蚕核型多角体病毒(BmNPV)orf126基因(Bm126)在感染宿主中的作用机制,采用PCR方法从BmNPV陕西株中克隆了切除N端23个氨基酸(预测信号肽序列)的Bm126基因,并将其和谷胱甘肽巯基转移酶(GST)融合后在大肠杆菌中进行了表达.利用纯化的BM126重组蛋白与几丁质进行体外结合实验,Western blot分析没有检测到BM126重组蛋白与几丁质结合的复合物,推测原核表达的重组BM126融合蛋白在体外不能与几丁质结合.
為瞭研究傢蠶覈型多角體病毒(BmNPV)orf126基因(Bm126)在感染宿主中的作用機製,採用PCR方法從BmNPV陝西株中剋隆瞭切除N耑23箇氨基痠(預測信號肽序列)的Bm126基因,併將其和穀胱甘肽巰基轉移酶(GST)融閤後在大腸桿菌中進行瞭錶達.利用純化的BM126重組蛋白與幾丁質進行體外結閤實驗,Western blot分析沒有檢測到BM126重組蛋白與幾丁質結閤的複閤物,推測原覈錶達的重組BM126融閤蛋白在體外不能與幾丁質結閤.
위료연구가잠핵형다각체병독(BmNPV)orf126기인(Bm126)재감염숙주중적작용궤제,채용PCR방법종BmNPV협서주중극륭료절제N단23개안기산(예측신호태서렬)적Bm126기인,병장기화곡광감태구기전이매(GST)융합후재대장간균중진행료표체.이용순화적BM126중조단백여궤정질진행체외결합실험,Western blot분석몰유검측도BM126중조단백여궤정질결합적복합물,추측원핵표체적중조BM126융합단백재체외불능여궤정질결합.
In order to study the functionaI mechanism of orf126 gene(Bm126)of Bombyx mori nucleopolyhedrovirus (BmNPV)during infecting the host,the Bm126 gene excising the N-terminal 23 amino acids of predicted signal peptide sequence was amplified by PCR from BmNPV Shaanxi isolate and was fused with glutathione S-transferase(GST)gene for expression in E,coli.in vitro binding assay was conducted for the purified recombinant BM126 protein with chitin.Western blot analysis did not identify any complex formed between the recombinant BM126 protein and chitin.sugges-ting that the prokaryotic expressed recombinant BM126 fusion protein iS unable to bind chitin in vitro.
