39 results about "Double mutation" patented technology

The invention is based on the reaction of recombinagenic oligonucleotides in a cell-free system containing a cytoplasmic cell extract and a test duplex DNA on a plasmid. The reaction specifically converts a mutant kanr gene to recover the resistant phenotype in transformed MutS, RecA deficient bacteria and allows for the rapid and quantitative comparison of recombinagenic oligonucleobases. Using this system a type of Duplex Mutational Vector termed a Heteroduplex Mutational Vector, was shown to be more active in than the types of mutational vectors heretofore tested. Further improvements in activity were obtained by replacement of a tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2'-O-methyl-uridine, to link the two strands of the Duplex Mutational Vector and removal of the DNA-containing intervening segment. The claims concern Duplex Mutational Vectors that contain the above improvements. In an alternative embodiment the claims concern a reaction mixture containing a recombinagenic oligonucleobase, a cell-free enzyme mixture and a duplex DNA containing a target sequence. In yet an alternative embodiment, the invention concerns the use of such mixture to test improvements in recombinagenic oligonucleobases, as well as to test the effects of compounds on the activity of the cell-free enzyme mixture and also to make specific changes in the target DNA sequence.

Modified polypeptides based on the botulinum neurotoxin A heavy chain containing the double mutation Trp-Tyr->Leu-Ser in the ganglioside binding motif Ser-X-Trp-Tyr do not bind polysialogangliosides and nerve endings. The polypeptides are useful in the preparation of nontoxic vaccines against the effects of C. botulinum infection. The modified polypeptides are also useful as vehicles for the transepithelial delivery of diagnostic and therapeutic entities, through formation of conjugates between the polypeptides and the diagnostic or therapeutic entities.

The invention relates to a rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application, and belongs to the technical field of biology. The gene mALS3 is obtained by introducing a new mutant site into the ALS3 gene sequence of the herbicide-resistant rape M342, and the site is positioned at the 560th site of the ALS3 gene; the original gene is alanine with a C alkaline group at the 187th site, the C alkaline group is mutated into a T alkaline group, and valine is formed at the 187th site. The rape mALS3 gene with double mutation sites A187V and W556L is obtained through an in-vitro site-specific mutation technique. The double mutation sites are not reported in the literatures of the rape genome. After preliminary functional verification of transgenic arabidopsis, the rape mALS3 gene with double mutation sites has the advantages that the obvious resistance on multiple ALS inhibitor herbicides, such as imidazolamide, tribenuron-methyl, difluorosulfonamide and bispyribac-sodium, is realized; the resistance gene can be used for the culturing of herbicide-resistant crops.

The invention discloses application of a UB2 / UB3 gene in regulation and control of corn multi-spike development, and particularly relates to application of UB2 and UB3 gene double mutation in regulation and control of corn multi-spike development. According to the invention, gene editing is carried out on two targets UB2 and UB3 in corn by using a CRISPR / Cas9 transgenic technology, so that the target site of the UB2 gene has deletion of a basic group to cause frameshift mutation and deletion of large fragments of the UB3 gene. Results show that the tassel branch number of UB2 and UB3 double mutant plants is reduced, the increase of the spike number of the UB2 and UB3 double mutants is observed for the first time, new female spikes grow on spike stalks of main spikes, the multi-spike property is shown, auxiliary meristem at the lower part is also derepressed to be developed into female spikes, and phenotypes with increased aerial roots are accompanied; and it shows that after the UB2 gene and the UB3 gene are knocked out, a corn mutant plant can show a multi-spike phenotype. The invention provides a theoretical basis for genetic improvement of the corn multi-spike property has a large application prospect.

We disclose a cell having double mutations of the hERG gene that lead to charge reversal amino acid substitutions at residues 466 and 534 of the wild type Kv11.1 channel protein. These double charge reversal mutations result in cells having constitutively open Kv11.1 channels. Such cells could be used in a method of testing development-stage drugs and other compounds for Kv11.1 channel block activity.

Disclosed is a new use of ibrutinib (PCI-32765). In particular, the present invention has found that ibrutinib can treat non-small cell lung cancer carrying EGFR T790M mutations and / or EGFR L858R mutations and / or EGFR delE746_A750 mutations, especially non-small cell lung cancer carrying EGFR L858R and EGFR T790M double mutations.

The invention relates to a double mutation detection method and a reagent box for a hepatitis B virus DNA A1762T-G1764A, in particular to a double mutation detection method for detecting the A1762T-G1764A, in the DNA sequence of the hepatitis B virus by utilizing a molecular beacon probe technology and a real time PCR method as well as a reagent box for finishing the detection. The method of the invention can specially and sensitively detect the double mutation hepatitis B virus DNA of the A1762T-G1764A in a clinic blood sample.

Compositions and methods are provided for diagnosis, prognosis and treatment of AR-related diseases, such as prostate diseases, such as prostate cancer, breast cancer, and many other diseases. In particular, a novel and clinically relevant mutation at position 876 of the androgen receptor (AR) has been identified. Drug sensitivity can be predicted and therapeutic regimens can be planned on the basis of the presence or absence of this mutation. Polypeptides comprising, antibodies to, and polynucleotides encoding the mutant AR can be used to identity novel treatments. A double mutation in AR at positions 741 and 877 is also shown to be useful for patient stratification.

The present invention provides a mutant of Perakine reductase and its application. The mutant is obtained by the following single mutation or combined double mutation of 126 and 127 in the amino acid sequence of wild-type Perakine reductase at position 126 or 127: H126G / R / I / C / P / K / N / W / A / Y, R127M / P / N / S / F, where " / " stands for "or". Based on the three-dimensional crystal structure and sequence comparison, the present invention adopts the point-saturation mutation technology to carry out molecular transformation on Perakine reductase, and screens to obtain the mutant capable of selectively reducing the enone double bond. The method of the present invention as a catalyst for selectively reducing enone double bonds has the advantages of simple reaction steps, mild reaction conditions, environmental friendliness, high catalytic efficiency, simple enzyme purification, good stability, and strong stereoselectivity. Bond reduction and the synthesis of related saturated ketone drugs have good application prospects.

The invention discloses a linoleate isomerase mutant and application thereof, and the linoleate isomerase mutant is obtained by mutation of 62th-site methionine of linoleate isomerase with an amino acid sequence as shown in SEQIDNO:1 into alanine or glycine, and mutation of 295th-site serine into alanine, leucine or valine. The enzyme activity for catalysis of linoleic acid for preparation of trans-10,cis-12 conjugated linoleic acid of the linoleate isomerase mutant can be greatly improved compared with that of a wild-type enzyme, especially during double mutation such as M62A / S295L, M62A / S295A, M62A / S295V, M62G / S295L, M62G / S295A and M62G / S295V, the enzyme activity can be increased by about 10 times, and the linoleate isomerase mutant has good prospects for industrial application.

The invention discloses a mutant for trehalose synthase as well as a preparation method and an application of the mutant and belongs to the fields of genetic engineering and enzyme engineering. The mutant is prepared by the following steps: mutating 289th bp glutamate near the active center of the trehalose synthase of Thermobifida fusca YX into glycine to obtain a mutant E289G; mutating 295th bp histidine into an asparagine mutant H295N; mutating 344th bp methionine into a lysine mutant M344K; and mutating 367th bp methionine into a leucine mutant M367L, and carrying out double mutations on the basis of the H295N to obtain mutants H295N / E289G, H295N / M344K, H295N / M367L and H295N / M344K / M367L. According to the mutant disclosed by the invention, although a substrate contains a certain quantity of glucose, the transformation efficiency of preparing trehalose by virtue of the trehalose synthase can not be greatly influenced still, and the mutant has very high industrial value.