6 results about "Homologous sequence" patented technology

The present invention relates to polynucleotide comprising a lncRNA gene according to SEQ ID No: 2, or according to a sequence having at least 50% sequence identity to SEQ ID No: 2, and an AP2c gene according to SEQ ID No: 3, or according to a sequence having at least 50% sequence identity to SEQ ID No: 3, which polynucleotide confers an increased density of type IV trichomes. The invention further relates to a plant comprising said polynucleotide and a SlASAT4 gene according to SEQ ID No. 21, or comprising a homologous sequence having at least 99.6% sequence identity to SEQ ID No. 21 wherein the presence of the polynucleotide leads to increased density of type IV trichomes and wherein the presence of said SlASAT4 gene leads to an increased ratio of S4:17 / S3:15 acyl sugars produced in glandular trichomes present on said plant when compared to a control plant lacking the polynucleotide and the SlASAT4 gene. Such a plant also was found to be resistant to Bemisia tabaci. The invention also discloses markers for the identification of the polynucleotide and the SlASAT4 gene and their use for the identification and development of Solanum lycopersicum plants. Also methods for identifying, producing and selecting of plants and seed are disclosed.

Provided are methods comprising expressing in a recombinant cell comprising a Cas gene a recombinant error-prone reverse transcriptase (RT) and a recombinant spacer RNA comprising a target sequence for mutagenesis of a DNA sequence in the Cas gene; making a mutagenized cDNA polynucleotide homologous to the DNA sequence in the recombinant cell; expressing a recombinant recombineering system in the recombinant cell; and recombining the mutagenized cDNA with the homologous DNA sequence of the Cas gene in the recombinant cell. Also provided are recombinant cells comprising recombinant coding sequences for a recombinant Cas protein, recombinant error-prone reverse transcriptase (RT), recombinant spacer RNA comprising the target sequence, and recombinant recombineering system.

The invention relates to the technical field of DNA cloning, and discloses a universal short-fragment DNA cloning method based on reverse PCR and template-free PCR and application of the universal short-fragment DNA cloning method, and the universal short-fragment DNA cloning method comprises the following cloning steps: taking an annular plasmid as a template, and adopting reverse PCR amplification to obtain a linearized vector; an upstream primer and a downstream primer are designed and synthesized, the 3'ends of the two primers have complementary overlapping regions, the 5 'ends of the two primers contain plasmid homologous sequences, and the two primers are subjected to thermal cycle self-assembly and extension in a PCR reaction system without exogenous template DNA to form a blunt-end double-stranded DNA fragment; and purifying the blunt-end double-stranded DNA fragment and a linearized vector, connecting through homologous recombination, and transforming a host cell to obtain a recombinant plasmid containing a target gene. The universal cloning method for the short-fragment DNA does not depend on any restriction enzyme cutting site, and the vector is universal; meanwhile, an exogenous template DNA is not needed; time and cost are greatly reduced, and the method is suitable for construction of high-throughput sgRNA, epitope tags, promoter elements and the like.

The purpose of the present invention is to provide: a double-stranded RNA for inhibiting the expression of both human YAP1 and human WWTR1; and a pharmaceutical composition comprising the same. This double-stranded RNA (dsRNA) for inhibiting the expression of both human YAP1 and human WWTR1 includes a sense strand and an antisense strand. The antisense strand includes a region complementary to a homologous sequence in a human YAP1 transcription product or in a human WWTR1 transcription product. The homologous sequence is a nucleotide sequence formed of 17 nucleotides in the human YAP1 transcription product, or is a nucleotide sequence formed of 17 nucleotides in the human WWTR1 transcription product. When the two nucleotide sequences are aligned, at least 14 nucleotides in consecutive 17 nucleotides are identical. Each of the strands has a length of at least 15 nucleotides. The sense strand and the antisense strand form a double-stranded region.

The application provides a multi-copy plasmid of Rhodotorula mucilaginosa, Rhodotorula mucilaginosa transformation, a Rhodotorula mucilaginosa strain with efficient overexpression of a target gene and a construction method of the Rhodotorula mucilaginosa strain, and relates to the technical field of genetic engineering. The construction method of the plasmid comprises the following steps: screening geneticin as a resistance marker of wild Rhodotorula mucilaginosa; taking plasmid YEp352 as a template and P1-F / R as primers to amplify a linearized plasmid fragment; taking a DNA sequence of a geneticin resistance expression box with a homologous arm as a template and G1-F / R as primers to obtain a target gene fragment with a homologous sequence; assembling the linearized plasmid fragment and the target gene fragment with the homologous sequence, and then transforming into an E. coli competent cell; and finally, positive colony identification is performed. The multi-copy plasmid provided by the application has multiple cloning sites and a selection marker, so that the genome modification in the Rhodotorula mucilaginosa is more flexible and convenient, and the target gene can be stably and efficiently expressed.

The present invention relates to a modified WAK gene comprising a deletion and / or a substitution, and / or an insertion when compared to the wild-type sequence comprising SEQ ID No. 1 or compared to a homologous sequence having at least 80% sequence identity to SEQ ID No. 1, wherein said modified WAK gene leads to Corynespora cassiicola resistance in a plant. The function of the encoded WAK protein is reduced, changed or absent. The invention further relates to plants comprising the modified gene and showing resistance to a Corynespora cassiicola.