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orf6 - complementation test Wild type + pNBV1 Wild type + pNBV1 + orf6 gene

orf6 - complementation test Wild type + pNBV1 Wild type + pNBV1 + orf6 gene orf6 - mutant + pNBV1 orf6 - mutant + orf6 gene Complementation test 수행 (SMB-glucose, SMB-CO) Primer extension orf1-6 , cutR , cutB 에 대한 primer extension 및 RT-PCR 수행 FixAB cloning

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orf6 - complementation test Wild type + pNBV1 Wild type + pNBV1 + orf6 gene

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  1. orf6- complementation test Wild type + pNBV1 Wild type + pNBV1 + orf6 gene orf6-mutant + pNBV1 orf6-mutant + orf6 gene Complementation test 수행 (SMB-glucose, SMB-CO) Primer extension orf1-6, cutR, cutB에 대한 primer extension 및 RT-PCR 수행 FixAB cloning pGEM T-easy vector에 PCR product cloning fixA, fixB결과 확인

  2. 1Kb cutC cutA orf2 orf3 orf4 cutB orf5-1 cutR cutC cutA orf2 orf3 orf4 orf6 orf1 cutB orf5-3 5 : Primer 5F and 6R 1 : Primer 1F and BR 4 : Primer 4F and 5R 3: Primer 3F and 4R 2 : Primer 3F and 5R Primer 1F : orf1-cutB RT-F Primer BR : orf1-cutB RT-R Primer 3F : orf3del-F Primer 4R : orf3del-R Primer 4F : orf4del-F Primer 5R : orf5del-R Primer 5F : copy2-P3 Primer 6R : copy2-P2 RT-PCR cDNA(O) RT-PCR cDNA(X) PCR SM 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 : Primer 1F and BR : 61 ℃ 2 : Primer 3F and 5R : 61 ℃ 3 : Primer 3F and 4R : 61 ℃ 4 : Primer 4F and 5R : 61 ℃ 5 : Primer 5F and 6R : 61℃

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