Rapid method to identify expression vectors and tansgenic plants based on pBI121 plasmid
To screen rapidly positive clones, recombinant plant expression vectors constructed based on the binary vector pBI121, containing sense, antisense or RNA interference (RNAi) cassette respectively, were amplified by PCR with a universal primer pair designed according to the flank sequence of multiple...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2008-03-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.02.003 |
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| Summary: | To screen rapidly positive clones, recombinant plant expression vectors constructed based on the binary vector pBI121, containing sense, antisense or RNA interference (RNAi) cassette respectively, were amplified by PCR with a universal primer pair designed according to the flank sequence of multiple clone site in pBI121, and PCR products with predicted sizes were further verified by digestion with the restriction enzyme BamHI. The PCR amplification-digestion with BamHI method was used for screening transgenic plants carrying the recombinant pBI121 expression vector, and detecting results of the method were consistent with the traditional means for identification of transgenic plants. In addition, PCR products amplified from positive clones harboring recombinant expression vectors may be sequenced to precisely demonstrate whether interesting fragments were inserted into pBI121 in a right orientation and whether mutation existed in the inserted sequences. These results show that a rapid and effective method to identify the expression vectors and their transgenic plants containing pBI121 plasmid was developed. |
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| ISSN: | 1008-9209 2097-5155 |