Beet M14 Strain Identification And Flowering Of The Fie Gene Expression Cdna Library Construction

FIE gene, which was first isolated in Arabidopsis thaliana, controls the development of fertilization-independent endosperm, and it might be one of components in apomixis. According to the conservation of FIE, We use methods of conservative sequence and attempt to find homological genes in Beta vulgaris M14.PCR primers were designed according to FIE gene sequence,than amplified FIE specific fragment of total DNA in Arabidopsis thaliana, linked to pGEM-T Easy Vector and transformed E.coli JM109. Restriction analysis and sequence analysis have proved that the colne product is FIE specific fragment of Arabidopsis thaliana. Comparing with FIE gene of Arabidopsis thaliana shared 99.8% homology at nucleotide level. It can be used as probes.By using FIE specific fragment as template,probes were labeled by digoxigenin(DIG). The results of Southern hybridization with genomic DNA in Beta vulgaris M14 suggest that M14 has homological genes of FIE. Total RNA was extracted from the bud of Beta vulgaris M14 followed by purification of mRNA, synthesis of the first strand and the second strand cDNA and obtained FIE specific fragment of 406bp of Beta vulgaris M14 by using RT-PCR. In order to obtain the gene of full length, the cDNA Library of M14 bud was constructed through highly efficient and simplified method. The resultant cDNA library will be used for the screeing of target genes of Beta vulgaris M14.