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Screening method of genome BAC homologous region of 12 rice species

A screening method, genome technology, applied in the field of molecular biology

Inactive Publication Date: 2015-12-09
HENAN INST OF SCI & TECH
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  • Claims
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Problems solved by technology

against MOC1 And its BAC homology region is screened by two rounds of southern hybridization with a specially designed scheme MOC1 The homologous region BAC of the 12 species of Oryza where it is located has not yet been specifically reported

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  • Screening method of genome BAC homologous region of 12 rice species
  • Screening method of genome BAC homologous region of 12 rice species
  • Screening method of genome BAC homologous region of 12 rice species

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Experimental program
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Embodiment Construction

[0035] 1 Southern probe preparation.

[0036] 1.1 Probe type.

[0037]There are two types, one is "anchor probe", which is used for the first round of screening of the wild rice BAC library; the other is "limited probe", which is used for further screening of positive clones identified in the first round of screening. tiller control gene MOC1 , the latter being MOC1 Upstream 2 genes (distance from MOC1 about 42kb, 53kb respectively) probes and 3 downstream genes (distance from MOC1 about 42kb, 52kb, and 62kb respectively) probes.

[0038] 1.2 Probe PCR amplification.

[0039] Take the japonica rice Nipponbare ( Oryza sativa L. ssp. japonica cv.Nipponbare) Genomic DNA was used as template for PCR amplification. The primer sequences are listed in Table 1.

[0040] "Anchor Probe" MOC1 Because of its high GC content (>75%), GC buffer (GCbufferⅠ, containing 5mmol / LMg2+) was used for amplification. The system is: LA Taq Enzyme, 0.3μL; 2×GCbufferⅠ, 12.5μL; dNTP, 3μ...

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Abstract

The invention discloses a screening method of genome BAC homologous region of 12 rice species. Developing a comparative study on a genome BAC homologous region, where MOC1 is, of 12 rice species with close genetic relationship, including diploid, tetraploid and other different genome types, is a powerful means for learning about evolution of the rice MOC1 genome region and can provide a certain theoretic reference for breeding application of import trait genes. As for MOC1 and the BAC homologous region, where MOC1 is, of the 12 rice species, a specially designed scheme is adopted for twice southern hybridization to realize quick and effective screening of the BAC homologous region, where MOC1 is, of the 12 rice species. The screening method provided by the invention solves the problem of how to quickly and effectively screen genomes BAC which are long enough as far as possible of homologous sequence region overlaps from genome BAC libraries of wild sibling species of the whole different rice genome types.

Description

technical field [0001] The invention relates to a genome BAC (bacterial artificial chromosome) homology region screening method in the technical field of molecular biology, in particular to a method for screening genome BAC homology regions by performing two rounds of southern hybridization through a specially designed scheme. Background technique [0002] Genome comparative studies on multiple plant species with close kinship including diploid, tetraploid and other genome types are a powerful means to understand the development and evolution of plant genes and genomes. The comparison of the entire genomes of multiple species can give a more comprehensive understanding of the analyzed species, but due to the huge amount of information in the whole genome, the analysis of some of the more important gene loci is not detailed. The green revolution in agricultural production, including rice and wheat, was triggered by the utilization of a few key genes. For example, the developm...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6816C12Q2537/149
Inventor 张胜利李东方胡海燕郝薇薇赵俊杰贺杰
Owner HENAN INST OF SCI & TECH