Protein encoding sequence of cucumber side branch inhibition gene cls

A coding sequence, a technology for inhibiting genes, applied in the coding sequence field in the field of genetic engineering technology

Inactive Publication Date: 2008-08-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the analysis to prior art document, although " Proc.Natl.Acad.Sci.USA Journal of the National Academy of Sciences ", 1999,1,96:290-295 published article " The lateral suppressor (Ls) gene of tomato encodes a The new member of the VHIID protein family "The protein encoded by the tomato collateral suppressor gene Ls is a new member of the VHIID family" elaborated on the function of the tomato collateral suppressor gene Ls; "Theor Appl Genet" in 2003, 107: 875 The article "Comparative analysis of response to phenotypic and marker-assisted selection for multiple lateral branching incucumber (Cucumis sativus L.)" published by -883 carried out QTLs for multiple lateral branching traits Mapped, but not isolated genes

Method used

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  • Protein encoding sequence of cucumber side branch inhibition gene cls
  • Protein encoding sequence of cucumber side branch inhibition gene cls
  • Protein encoding sequence of cucumber side branch inhibition gene cls

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Cloning of Example 1 Cucumber Collateral Suppressor Gene

[0036] 1. Tissue separation (isolation)

[0037] Cucumber s06 is a European greenhouse variety collected in our laboratory. The cucumber was germinated at 28°C for 24 hours, and then sown in plug trays. After the two cotyledons of the cucumber were completely flattened, it took about a week to prepare for DNA or RNA extraction.

[0038] 2. RNA isolation (RNA isolation)

[0039] Take part of the tissue, grind it with a mortar, add it to a 1.5mL EP tube filled with lysate, shake it fully, and then transfer it into a glass homogenizer. After homogenization, transfer to 1.5mL EP tube, and extract total RNA (TRIzol Reagents, GIBCO BRL, USA). The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0040] 3. Cloning of Full-length cDNA

[0041] According to the nucleotide conserved sequence of Arabidopsis, tomato, and ...

Embodiment 2

[0052] Example 2 Sequence information and homology analysis of genes related to cucumber collateral suppression

[0053] The full-length cDNA of the new cucumber collateral suppressor gene in this example is 2175bp (SEQ ID NO.3), wherein the open reading frame is located at nucleotides 491-1666 (1175 nucleotides). According to the full-length cDNA, the amino acid sequence of the cucumber collateral suppressor gene is deduced, with a total of 391 amino acid residues, and the detailed sequence is shown in SEQ ID NO.3.

[0054] The full-length cDNA sequence related to the cucumber collateral suppressor gene and its encoded protein were compared in the Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDStranslations+PDB+SwissProt+Superdate+PIR databases using the BLAST program. Protein homology search, it was found that it has homology with tomato collateral suppressor gene (AF098674) only in some nucleotide segments (attached table 2), and has 67% identity at the ami...

Embodiment 3

[0067] The subcellular localization of embodiment 3cls gene

[0068] The functions of GRAS family protein members are transcription factors, based on the discovery that some GRAS members, such as RGA and SLR1, have nuclear localization signals. However, analysis with molecular biology software (pSORT, http / / psort.nibb.ac.jp) showed that CLS has no transmembrane region. In order to determine whether CLS is localized in the nucleus, a fusion protein containing green fluorescent protein GFP was constructed: CLS -GFP, constructed into the chronological expression vector PA-7. Bombarded into the epidermis of the onion with a gene gun, cultured in the dark at 24°C for 36 hours, and then observed with a laser confocal microscope. Results The green fluorescent signal was mainly detected in the nucleus, indicating that CLS was localized in the nucleus, presumably a transcription factor.

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Abstract

The invention relates to a coding sequence of green cucumber collateral inhibition gene CLS in genetic engineering technology field. The coding sequence comprises coding polypeptides nucleotide sequence with cucumber collateral inhibition albumen activity, and the nucleotide sequence and 491-1666 bite nucleotide sequence in SEQ ID NO.3 have at least 701mology. The opening reading frame of coding sequence is 1175 bases from 491th to 1666th. The invention can stimulate forming of lateral bud anlage and is used for alimentary crops, floral plants, economic crop and oil crop to increase the number of branch stem thereby raise productivity.

Description

technical field [0001] The present invention relates to a coding sequence in the technical field of genetic engineering, in particular to a protein coding sequence of cucumber collateral suppressor gene cls. Background technique [0002] Plant branch development plays an important role in plant morphogenesis. Crop branching is one of the important agronomic traits affecting crop yield. In the process of domestication of maize, the purpose of increasing yield can be achieved by reducing branching. Too many or too few tillers in rice are not conducive to the increase of yield. For cucumbers, the yield is also closely related to the number of side branches. So far, cucumber collaterals have been studied as a quantitative trait, and several QTLs related to collateral development have been mapped. But the gene has never been isolated, let alone studied in detail. However, multiple genes related to lateral branch development have been isolated in maize, rice, Arabidopsis, tomat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/79C12N1/19C12N5/10C07K14/415C12Q1/68A01H1/00A01H5/00
Inventor 原丽华蔡润朱立煌姚丹青张弛
Owner SHANGHAI JIAO TONG UNIV
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