LdMNPV-NEFU isolate collected from the forestry farm of Northeast Forestry University was purified and the genomic DNA of LdMNPV was extracted. The LdMNPV polyhedrin gene was cloned by PCR. The results showed that the...LdMNPV-NEFU isolate collected from the forestry farm of Northeast Forestry University was purified and the genomic DNA of LdMNPV was extracted. The LdMNPV polyhedrin gene was cloned by PCR. The results showed that the sequence was an open reading frame (ORF) of 735bp capable of encoding 245 amino acids. The polyhedrin gene sequences of the LdMNPV-NEFU isolate and a Canada strain, LdMNPV-G differed in 5 bases. The polyhedrin gene of the LdMNPV-NEFU isolate contained C, G, T, C and G at 54, 109,379, 508 and 701 sites from the start codon, but the LdMNPV-G isolate contained G, C, C, T and T at the corresponding sites respectively. The same amino acids were encoded by the two ORF sequences, with the exception that Asp and His are encoded by GAC on the polyhedrin gene sequence of the LdMNPV-NEFU isolate and by CAC in the LdMNPV-G isolate. The LdMNPV polyhedrin gene was expressed in E.coli BL21 (DE3) by the pT7-7 plasmid vector.展开更多
The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as t...The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as the probe.In order to sequence the EpNPV polyhedrin gene,the Bam H I G fragment was cloned into pTZ19R vector.Then the fragment was digested by Xba I? Hin d III,and subcloned into pTZ19R.展开更多
文摘LdMNPV-NEFU isolate collected from the forestry farm of Northeast Forestry University was purified and the genomic DNA of LdMNPV was extracted. The LdMNPV polyhedrin gene was cloned by PCR. The results showed that the sequence was an open reading frame (ORF) of 735bp capable of encoding 245 amino acids. The polyhedrin gene sequences of the LdMNPV-NEFU isolate and a Canada strain, LdMNPV-G differed in 5 bases. The polyhedrin gene of the LdMNPV-NEFU isolate contained C, G, T, C and G at 54, 109,379, 508 and 701 sites from the start codon, but the LdMNPV-G isolate contained G, C, C, T and T at the corresponding sites respectively. The same amino acids were encoded by the two ORF sequences, with the exception that Asp and His are encoded by GAC on the polyhedrin gene sequence of the LdMNPV-NEFU isolate and by CAC in the LdMNPV-G isolate. The LdMNPV polyhedrin gene was expressed in E.coli BL21 (DE3) by the pT7-7 plasmid vector.
文摘The polyhedrin gene of EpNPV was located on Hin d III E, Xba I B, Eco RI J, Kpn I A, Pst I F, Bam H I G fragments under stringent condition(68℃,in water),by using the DIG labeled Bam H I F fragment of AcMNPV DNA as the probe.In order to sequence the EpNPV polyhedrin gene,the Bam H I G fragment was cloned into pTZ19R vector.Then the fragment was digested by Xba I? Hin d III,and subcloned into pTZ19R.
