CONSTRUCTION OF PLANT TRANSFORMATION VECTOR: SUB-CLONING OF PIGEONPEA LECTIN (CCL) GENE UNDER rolC PROMOTER
KrishiKosh
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Title |
CONSTRUCTION OF PLANT TRANSFORMATION VECTOR: SUB-CLONING OF PIGEONPEA LECTIN (CCL) GENE UNDER rolC PROMOTER
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Creator |
DEEPAK VISHWANATH PAWAR
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Contributor |
Rekha Kansal
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Subject |
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Description |
T-8621
The 928bp rolC promoter (accession number DQ-160187) was PCR amplified from the bacterial culture of rolC promoter cloned in pUC29 vector using promoter specific primers containing restriction enzyme sites of XbaI in forward and EcoRI in reverse. The amplified product was then restricted with restriction enzymes XbaI & EcoRI and ligated with pORE O4 binary vector digested with the same restriction enzymes. 828bp Pigeonpea lectin (CCL) was restricted from the bacterial culture of pigeonpea lectin gene cloned in pGEMT vector with the restriction enzyme NotI and ligated with the promoter construct such that CCL gene will be cloned downstream of rolC promoter in pORE O4 binary vector. The clones were confirmed by restriction digestion and colony PCR, The gene construct was mobilized into Agrobacterium tumefacience GV-3101, which will be used for transformation of Brassica juncea to express pigeon pea lectin protein in phloem sap which is desirable in case of Hemipteran (phloem sap sucking) pests like aphids. Keywords: aphid, pigeonpea lectin (CCL) gene, rolC pr |
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Date |
2016-09-21T16:34:46Z
2016-09-21T16:34:46Z 2012 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/78183
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Format |
application/pdf
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Publisher |
IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY
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