Saccharum spontaneum retrotransposon sequence and identification method thereof

A technology of retrotransposon and identification method, which is applied in the field of retrotransposon sequence and its identification, which can solve the problem of small shape, poor effect, and lack of rapid and accurate identification of the chromosome Probe and other problems, to achieve the effect of good primer specificity, single and bright band

Active Publication Date: 2019-08-30
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Since Saccharum species are mostly highly heterozygous polyploid plants, their chromosomes are large in number, small in shape and similar in shape, and there is still no specific probe that c

Method used

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  • Saccharum spontaneum retrotransposon sequence and identification method thereof
  • Saccharum spontaneum retrotransposon sequence and identification method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1: According to the genome clustering data analysis of SES208 and tropical species LA Purple, the Sequence Retrotransposon sequence of Sequence 1-Cluster168Contig15, Sequence 2-Cluster56Contig54, Sequence 3-Cluster100Contig20, Sequence 4-Cluster38Contig50, the nucleotide sequences of which are respectively shown in SEQ ID NO.1-4. The molecular marker primer sequences corresponding to the sequence 1-Cluster168Contig15, the sequence 2-Cluster56Contig54, the sequence 3-Cluster100Contig20, and the sequence 4-Cluster38Contig50 are shown in SEQ ID NO.5-12.

[0036] Table 1 Amplification primers for reverse transcription of transposon sequence

[0037]

[0038] Primers were designed for retrotransposon sequences using Primer Premier 5.0.

[0039] Perform PCR amplification on the above four cut-hand-closed retrotransposon sequences. The PCR amplification conditions are: pre-denaturation at 95°C for 3 minutes; denaturation at 98°C for 30 seconds, annealing and extens...

Embodiment 2

[0040] Example 2: Identification by fluorescence in situ hybridization (FISH)

[0041]Fluorescence in situ hybridization is a technique for detecting the distribution of target sequences on chromosomes. The detection results of this method are true and reliable. It is not only suitable for the identification of single-copy genes on chromosomes, but also for the identification of multiple copies of repetitive sequences on chromosomes. Identification studies.

[0042] The fluorescence in situ hybridization identification of the cutshoumi retrotransposon sequence includes the following steps:

[0043] (1) Take the vigorously growing SES208 and LA Purple root tips respectively, and put them into 8-hydroxyquinoline solution at room temperature for pretreatment;

[0044] (2) cutting off the root cap and the elongation zone, leaving the root tip meristem, and using cellulase and pectinase to enzymatically remove the root tip meristem cells at 37°C;

[0045] (3) Aspirate the root ti...

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Abstract

The invention discloses a saccharum spontaneum retrotransposon sequence and an identification method of the saccharum spontaneum retrotransposon sequence. Four saccharum spontaneum retrotransposon sequences are screened out using cluster analysis of genome data of saccharum spontaneum and a tropical species; primers are designed separately according to the sequences for amplification to form a full-length sequence; then, a probe is prepared; fluorescence in situ hybridization (FISH) identification is performed on metaphase chromosomes of the saccharum spontaneum and the tropical species; and aresult shows that the four retrotransposon sequences generate clear and bright signals on the saccharum spontaneum chromosomes. The retrotransposon sequence can be directly used for specific identification of the saccharum spontaneum chromosomes, provides more accurate information for blood relationship identification of the saccharum spontaneum in cultivated sugarcane varieties, and lays a foundation for chromosome engineering breeding of sugarcane.

Description

technical field [0001] The invention relates to the fields of bioinformatics and molecular cytogenetics, in particular to the reverse transcription transposon sequence and identification method of the cut hand secret. Background technique [0002] The content of repetitive sequences is one of the most important factors affecting the genome size of plants. In general, the greater the proportion of repetitive sequences in a plant, the larger its genome. For example, the Arabidopsis genome is relatively small, only 121Mb, and its repetitive sequences account for about 25%; the up to 17Gb wheat genome contains about 90% repetitive sequences. Retrotransposons are a type of repetitive sequences that widely exist in plant genomes. They use RNA as an intermediate to perform reverse transcription through their own encoded reverse transcriptase, and generate extrachromosomal DNA inserted into new target sites in the genome. , moving in a "copy-paste" manner in the genome, eventually...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6841
CPCC12Q1/6841C12Q2563/107
Inventor 王凯黄永吉韩金磊闫天盈
Owner FUJIAN AGRI & FORESTRY UNIV
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