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The beet M14 monosomic addition lines the additional chromosome BIBAC chip development and application

Author: DengDaiQian
Tutor: KangChuanHong
School: Heilongjiang University
Course: Biochemistry and Molecular Biology
Keywords: BIBAC Gene chip Apomixes M14 sugar beet Bacterial artificial chromosome (BAC)
CLC: S566.3
Type: Master's thesis
Year: 2006
Downloads: 45
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Abstract


M14 beet is a monosomic addition line of Beta corolliflora Zoss which constitutes of the normal 18 B.vulgaris chromosomes plus Beta corolliflora chromosome 9. According as the investigation we found that the transmission frequency of M14 up to 97% in successive four generations. The apomixis type of M14 is butterfly-beard, autonomous endosperm genesis but pollen stimulation is needed. We believed the gene(s) controlling apomixis may be located on the alien chromosome 9 from Beta corolliflora Zoss.In order to research the putative apomixes-controlling gene(s) of M14 beet, Doctor XiaohuaFang and adjunct professor HongbinZhang constructed a plant-transformation- competent binary BAC library from the genomic DNA of M14, the library consists of 49,920 clones and the average size of insert is 127kb. The total insert fragments of the library are 7.5 times larger than the genome of M14 and contain more than 99% genes of genome of M14. We used pBC1054 probe of the chromosome Beta corolliflora genome-specific dispersed repetitive DNA sequences to isolate BAC clones derived for the alien chromosome in the library. In result a total of 2,377 positive clones were obtained and arrayed into a sublibary specific for Beta corolliflora chromosome 9, these clones represent 3. 8 times of plus Beta corolliflora chromosome 9 equivalents.We have improved the alkaline-abruption method to isolate large BACplasmid and the method to attach BAC DNA convalent to modified surface ofglass. With these technologies we constructed the BAC chip with 2, 377 BACDNA from Beta corolliflora chromosome 9 in the array. The

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CLC: > Agricultural Sciences > Crop > Economic crops > Sugar crop > Beets ( sweet carrots)
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