Plants having enhanced yield-related traits and a method formaking the same
A yield-related trait, extensin technology, applied in the field of molecular biology
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Embodiment 1
[0905] Example 1: Identification of homologues of ERLK protein SEQ ID NO: 2 in Arabidopsis, rice and other plant species
[0906] Use database sequence search tools, such as Basic Local Alignment Tool (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402) in the country Among those sequences maintained in the Entrez Nucleotide Database of the Biotechnology Information Center, sequences related to the nucleic acid sequence used in the method of the present invention (full-length cDNA, EST or genome) were identified. This program is generally used to find regions of local similarity between sequences by comparing nucleic acid sequences or polypeptide sequences with sequence databases and calculating the statistical significance of matches. The TBLASTN algorithm is used for the polypeptide encoded by the nucleic acid of the present invention, and default settings and filtering are adopted to ignore low-complexity seque...
Embodiment 2
[0911] Example 2: Determination of global similarity and identity between the kinase domains of ERLK proteins
[0912] The percent similarity and percent identity between the kinase domains of ERLK proteins use MatGAT (Matrix Global Alignment Tool) software (BMC Bioinformatics.2003 4:29. MatGAT: Use protein sequence or DNA sequence to generate similarity / identity matrix The application. Campanella JJ, Bitincka L, Smalley J; software owned by Ledion Bitincka) to determine. MatGAT software generates similarity / identity matrices for DNA sequences or protein sequences without pre-alignment of data. Using the Myers and Miller global alignment algorithm (gap opening penalty of 12 and gap extension penalty of 2), the program performs a series of paired alignments, using, for example, Blosum 62 (for peptides) to calculate similarity and identity and then set the results In the distance matrix. Sequence similarity is shown in the lower half of the dividing line, and sequence identity is sh...
Embodiment 3
[0920] Example 3: Cloning of the nucleic acid sequence used in the method of the present invention
[0921]DNA manipulation: Unless otherwise stated, recombinant DNA technology is based on (Sambrook (2001) Molecular Cloning: a laboratory manual, third edition Cold Spring Harbor Laboratory Press, CSH, New York) or Ausubel et al. (1994), Current Protocols in Molecular Biology, Current Protocols 1 and 2 are described in standard methods. Standard materials and methods for plant molecular work are described in Plant Molecular Biology Labfax (1993) by R.D.D. Croy published by BIOS Scientific Publications Ltd (UK) and Blackwell Scientific Publications (UK).
[0922] The nucleic acid sequence used in the method of the present invention is amplified by PCR using a customized Arabidopsis thaliana seedling cDNA library (in PCMV Sport 6.0; Invitrogen, Paisley, UK) as a template. PCR was performed using Hifi Taq DNA polymerase under standard conditions using 200 ng template in 50 μl PCR mix. ...
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