Research Of Cloning The Helicase Gene From The Banana And Transforming Tomato

1) Cloning the helicase gene from the bananaIn this research, gradient PCR technique was used to amplify the object gene in cDNA library of banana. A 5'fragment was obtained, whose length was 804bp. After integrating it with the known fragment of the object gene, a cDNA sequence of object gene whose length was l,473bp was gained. The result of homologous analysis of the cDNA fragment and amino acids sequence using NCBI-Blast showed that it was helicase gene family. The result of Southern blotting indicated that it came from banana and only low copy.2) Antisense gene fragment transformed to tomatoThrough the construction antisense gene of plant expression vector, the agrobacterium-mediated method was used to transform the cotyledon of tomato. The screen of Kanamycin has got twenty plants. Seven of them, which had variation, were chosen to go on Southern blot. Five of them were transgenic plants. Compared with the control, there was too much mutation in morphology and pathology. This conclusion coincided with our prediction. Analysis of genetic phenotype of the transgenic plants shed light on understanding the function of the helicase gene.