In From Beets M14 Of Beta Corolliflora Cdna Library Screening Apomixis Candidate Gene

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, then amplified FIE specific fragment of 406bp in Beta vulgaris M14 by using RT-PCR. Sequence analysis suggest that the FIE specific fragment of 406bp codes 135 amino acid. Comparing with FIE gene of Arabidopsis thaliana, the FIE specific fragment of 406bp in Beta vulgaris M14 shared 82.407% homology at nucleotide acid level and 83.704% homology at amino acid level.Using the total RNA of bud, stem, root, leaf in Beta vulgaris M14 and bud in Beta.vulgaris L. as template, we designed the RT-PCR primer according to FIE specific fragment of Beta Beta Vulgaris M14 and then synthesized double cDNA. The result is that we amplified FIE specific fragment of 406bp in all tissues of Beta vulgaris M14 and in the bud of Beta.vulgaris L by using RT-PCR .Then we linked the fragment to pGEM-T Easy Vector and transformed E.coli JM109.Restriction analysis have proved that the clone product is FIE specific fragment of Beta vulgaris M14.So we know that the FIE specific fragment of 406bp exists in all the tissues of Beta vulgaris M14 and in the bud of Beta.vulgaris L. The conclusion is that the FIE specific fragment of 406bp hasn't the character of tissue-specific and matierial-specific.In order to obtain the gene of full length, the cDNA library of M14 bud was constructed through highly efficient and simple method. The probeof FIE specific fragment of 406bp was labeled with DIG and we take the first step in screening the target gene of FIE by phagocyte hybridization.