Potato virus Y (PVY) N coat protein (CP) coding sequence was cloned into a plant expression vector pMON316 under the CaMV 35S promoter. Leaf discs of potato (Solanum tuberosum) were used to Agrobacterium mediated gene...Potato virus Y (PVY) N coat protein (CP) coding sequence was cloned into a plant expression vector pMON316 under the CaMV 35S promoter. Leaf discs of potato (Solanum tuberosum) were used to Agrobacterium mediated gene transfer. A large number of regenerated putative transgenic plants were obtained based on kanamycin resistance. Using total DNA purified from transgenic plants as templates and two oligonucleotides synthesized from 5′ and 3′ of the PVY coat protein gene as primers, the authors carried out polymerase chain reaction(PCR) to check the presence of this gene and obtained a 0.8 kb specific DNA fragment after 35 cycles of amplification. Southern blot indicated that the PCR product was indeed PVY CP gene which had been integrated into the potato genome. Enzyme linked immunosorbent assay (ELISA) of our transgenic plants showed that CP gene was expressed in at least some transgenic potato plants.展开更多
Imported maize seeds were planted in a quarantine greenhouse and two seedlings with chlorotic mottle symptoms were observed.The ELISA results showed that the two seedlings reacted positively with antibody against Maiz...Imported maize seeds were planted in a quarantine greenhouse and two seedlings with chlorotic mottle symptoms were observed.The ELISA results showed that the two seedlings reacted positively with antibody against Maize chlorotic mottle virus(MCMV).The positive samples were further identified by RT-PCR method and the 711 bp size target bands were amplified specifically.The similarity range of nucleotide sequence of both RT-PCR product and six MCMV coat protein genes was 97%-99%.The amino acid sequence homology was 97.88%-99.89%.Based on the above results,the virus was identified as MCMV.It was the first time that MCMV was intercepted from imported maize seeds.展开更多
The coat protein gene and its 3′ noncoding region of a watermelon isolate of Cucumber green mottle mosaic virus (CGMMV-LN) in Liaoning, China was determined and compared with other isolates. The gene encoding the coa...The coat protein gene and its 3′ noncoding region of a watermelon isolate of Cucumber green mottle mosaic virus (CGMMV-LN) in Liaoning, China was determined and compared with other isolates. The gene encoding the coat protein of CGMMV-LN comprises of 486 nucleotides and encodes a putative protein of 161 amino acids. Sequence comparisons showed that the coat protein of this isolate was identical to that of seven CGMMV isolates infecting cucurbits in Europe and Asia. The 3′ noncoding region of CGMMV-LN consists of 176 nucleotides, which is same as that of CGMMV-KOM, CGMMV-KW and CGMMV–SH.展开更多
文摘Potato virus Y (PVY) N coat protein (CP) coding sequence was cloned into a plant expression vector pMON316 under the CaMV 35S promoter. Leaf discs of potato (Solanum tuberosum) were used to Agrobacterium mediated gene transfer. A large number of regenerated putative transgenic plants were obtained based on kanamycin resistance. Using total DNA purified from transgenic plants as templates and two oligonucleotides synthesized from 5′ and 3′ of the PVY coat protein gene as primers, the authors carried out polymerase chain reaction(PCR) to check the presence of this gene and obtained a 0.8 kb specific DNA fragment after 35 cycles of amplification. Southern blot indicated that the PCR product was indeed PVY CP gene which had been integrated into the potato genome. Enzyme linked immunosorbent assay (ELISA) of our transgenic plants showed that CP gene was expressed in at least some transgenic potato plants.
文摘Imported maize seeds were planted in a quarantine greenhouse and two seedlings with chlorotic mottle symptoms were observed.The ELISA results showed that the two seedlings reacted positively with antibody against Maize chlorotic mottle virus(MCMV).The positive samples were further identified by RT-PCR method and the 711 bp size target bands were amplified specifically.The similarity range of nucleotide sequence of both RT-PCR product and six MCMV coat protein genes was 97%-99%.The amino acid sequence homology was 97.88%-99.89%.Based on the above results,the virus was identified as MCMV.It was the first time that MCMV was intercepted from imported maize seeds.
文摘The coat protein gene and its 3′ noncoding region of a watermelon isolate of Cucumber green mottle mosaic virus (CGMMV-LN) in Liaoning, China was determined and compared with other isolates. The gene encoding the coat protein of CGMMV-LN comprises of 486 nucleotides and encodes a putative protein of 161 amino acids. Sequence comparisons showed that the coat protein of this isolate was identical to that of seven CGMMV isolates infecting cucurbits in Europe and Asia. The 3′ noncoding region of CGMMV-LN consists of 176 nucleotides, which is same as that of CGMMV-KOM, CGMMV-KW and CGMMV–SH.
