319 results about "Full length cdna" patented technology

Full-length cDNAs of plants and their uses are provided. Source plants are preferably monocot plants, more preferably poaceous plants, and most preferably rice. Vectors carrying said cDNAs and transformants containing said cDNAs or said vectors, transgenic plants containing said transformants, polypeptides encoded by said cDNAs are also provided. The full-length cDNA clones play important roles in the annotation of correct gene coding region, determination of exons and introns, comprehensive expression analysis on the transcription level and proteome analysis. Furthermore, full-length cDNA clones are industrially useful in producing plants having different properties from those of the wild type due to the inhibition of expression and functional suppression in plant bodies.

The present invention relates to a novel gene, Di12, that is differentially expressed as a 1.35 kb RNA in breast cancer tissues and cell lines, and in several normal tissues. The full length cDNA encodes a protein of 339 amino acids. Antibodies to the gene product were developed to investigate the expression of Di12 in breast cancer cell-lines and tumors. The Di12 protein was found in tissue sections of infiltrating ductal carcinomas (IDCs), but not in benign or normal breast specimens. Di12 wag also present in IDC-breast cancer patient sera, and its expression level increased markedly if IDC was accompanied by lymph node or distal metastases. As IDC constitutes ~70% of breast cancers seen clinically, the level of Di12 expression is useful for diseases diagnosis predicting disease progression and monitoring a therapeutic treatment.

A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3' and 5' ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.

Disclosed is a method for making full-length CDNA libraries, which is for making libraries of cDNAs having a length corresponding to a full-length of mRNAs and comprises the following steps of; binding a tag molecule to a diol structure present in 5' Cap (7MeGpppN) sites of mRNAs, forming RNA-DNA hybrids by reverse transcription using primers such as oligo dT and the mRNAs connected with the tag molecule as templates, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length of mRNAs from the RNA-DNA hybrids formed above by using function of the tag molecule. To obtain mRNA connected with a tag molecule, the diol structure present in 5' Cap site of mRNA is subjected to a ring-open reaction by oxidation with sodium periodate to form a dialdehyde and the dialdehyde is reacted with a tag molecule having a hydrazine terminus. According to the present invention, there are provided a novel method capable of efficiently labeling 5' Cap site and a method for making full-length cDNA libraries utilizing the labeling method.

The present invention relates to a novel gene, CaSm, that is highly expressed in cancer tissues and cell lines, especially pancreatic cancer. The full length cDNA of CaSm encodes a protein of 133 amino acids. CaSm contains the two Sm motifs found in the common snRNP proteins, with the greatest homology to the Sm G protein (60% similarity). The present invention further encompasses CaSm peptides, fusion proteins, host cell expression systems, antibodies to CaSm, antisense CaSM molecules, and compounds that modulate CaSm gene expression or CaSm activity. Antisense CaSm RNA is able to alter the transformed phenotype of pancreatic cancer cells by reducing their ability to form large colonies in soft agar when compared to untransfected cells. The present invention also encompasses methods for disease diagnosis, drug screening and the treatment of cancer.

A method of preparing normalized and/or subtracted cDNA; a method in which the cDNA that is normalized and/or subtracted is in the form of uncloned cDNA (cDNA tester); a method of preparing normalized and/or subtracted cDNA comprising the steps of: (a) preparing cDNA tester; (b) preparing normalization and/or subtraction RNA driver; (c) conducting normalization and/or subtraction in two steps in any order, or conducting normalization/subtraction as a single step and mixing the normalization/subtraction RNA driver with said cDNA tester; (d) adding an enzyme capable of cleaving single strand sites on RNA drivers nonspecifically bound to cDNA tester; (e) removing said single strand RNA driver cleaved in step d) from the tester and removing tester/driver hybrids; and (f) recovering the normalized and/or subtracted cDNA; and a method of efficiently preparing normalized and/or subtracted long-chain, full-coding, and full-length cDNA libraries are provided.

The invention discloses a plant flavonoid synthesis regulation gene and its application. CDNA obtained through the inverse transcription of the total RNA of Epimedium brevicornum Maxim blades is treated as a template and undergoes PCR to obtain a 233bp fragment, the 233bp fragment has a very high homology with flavonoid related R2R3-MYB gene, a full-length cDNA sequence is obtained through an RACE technology and is named EsMYBA1 gene, and the sequence of the EsMYBA1 gene is represented by SEQIDNO.1. The EsMYBA1 gene codes an R2R3-MYB transcription factor which has an about 50% homology with other MYB regulation genes for controlling the flavonoid synthesis approaches, and the improvement of the expression level of the EsMYBA1 gene through a genetic engineering technology promotes the synthesis and the accumulation of anthocyanidin.

PCT No. PCT/JP97/03992 Sec. 371 Date Jul. 16, 1999 Sec. 102(e) Date Jul. 16, 1999 PCT Filed Oct. 31, 1997 PCT Pub. No. WO98/20122 PCT Pub. Date May 14, 1998Disclose is a method for making full-length cDNA libraries, which is for making libraries of cDNAs having lengths corresponding to full lengths of mRNAs and comprises the following steps of; forming RNA-DNA hybrids by reverse transcription starting from primers using mRNAs as templates, chemically binding a tag molecule to a diol structure present in the 5' Cap (7MeGpppN) site of a mRNA which is forming a RNA-DNA hybrid, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length mRNA from the RNA-DNA hybrids formed above by using a function of the tag molecule. The present method is a method for preparing full-length cDNA libraries utilizing a method for labeling the 5' Cap site more efficiently than protein enzyme reactions, which is avoidable a decrease of a full-length cDNA synthesis efficiency caused by cleavage of mRNA, and can synthesize a full-length cDNA more efficiently.

The present invention relates to a novel gene, CaSm, that is highly expressed in cancer tissues and cell lines, especially pancreatic cancer. The full length cDNA of CaSm encodes a protein of 133 amino acids. The present invention further encompasses CaSm peptides, fusion proteins, host cell expression systems, antibodies to CaSm, antisense CaSm molecules, and compounds that modulate CaSm gene expression or CaSm activity. The present invention also encompasses methods for disease diagnosis, drug screening and the treatment of cancer. In particular, the combined use of a CaSm antagonist with a therapeutic agent to treat cancer is encompassed.

The invention belongs to a method for extensively screening scallop SNP in the technical field of the scallop DNA molecular genetic marker, which comprises the following steps that: solution D and phenol-chloroform are firstly used for extracting a plurality of Chlamys farreri RNA, and a ultraviolet specrophotometer is used for measuring the concentration of the mixture of solution D and the phenol-chloroform which are uniformly mixed; a scallop full-length cDNA sample is re-established and comprises the synthesis of a cDNA first chain and the synthesis and augmentation of a cDNA second chain; then homogenization of full-length cDNA is performed, and ultrasonic breaking is performed; then sequence measuring joints are connected, and terminal repair, joint connection and sample augmentation are included; finally a biological software is used for analyze the data to obtain an SNP marker; then an SNP marker to be screened is selected to design a primer; and DNA of individual scallop is extracted to be equivalently mixed to be used as a template, conventional colony sequence measuring is undertaken after the PCR augmentation, and the position point with the occurring frequency of single base difference being 10 percent or more is defined as an SNP marker. The method has safe and reliable principle, simple and controllable flow procedures, reliable running of large-scale screening, excellent effect and strong practicability.

A functional mutant of human plasminogen disclosed in the invention, respectively, is hPLG-delta K: human plasminogen protein Pro<544>-Asn<791> polypeptide; Pro<559> in RGD-hPLG-delta K: hPLG-delta K is mutated to Asp<559>; and Gly<560> in RHP-hPLG-delta K: hPLG-delta K polypeptide is mutated to His<560>. The invention also discloses a preparation method for the functional mutant. The product is obtained by using a plasmid containing a full-length cDNA sequence of human plasminogen as a template to carry out a PCR to construct a plasmid and using Pichia Pastoris expression. The functional mutant has a dual-function of fibrinolysis and inhibiting platelet aggregation or inhibiting fibrin monomer polymerization.

The invention relates to a virus antibody research and provides a method for constructing a hog-cholera virus infectious cDNA carrier having a molecule mark. The method comprises the following steps of: (1) diluting a hog-cholera live vaccine to 1 portion per milliliter as the experimental material, after extracting the head RNA, amplifying 6 corresponding cDNA fragments through RT-PCR section according to designed 6 pairs of primers, (2) leading in the corresponding molecule marks from F1 and F5 of cDNA fragments, (3) reforming plasmids required by overall length cDNA cloning, and (4) constructing the overall length cDNA carrier. A hog-cholera virus lapinized vaccine C stem infectious cDNA carrier pA-FL22 having a molecule mark can be acquired through the method of the invention. The permissive cells of RNA electro transferred hog-cholera virus acquired by the carrier can save the infectious progeny virus which can stably inherit the molecule mark led in the construction. By using the saving technique system of the infectious cDNA carrier and the progeny virus, the copy stem and the pathopoiesia reason of the virus can be deeply researched and the foundation for developing new marked vaccines can be settled.

The invention discloses a cDNA specific molecular tag and application thereof. The specific molecular tag comprises a universal primer sequence as shown in SEQ ID NO.1, a transposase joint sequence P7 as shown in SEQ ID NO.2, random sequences (N)n and a poly(T) sequence with 30 T alkalis, wherein N refers to the random sequences in a quantity of n, and a specific nucleotide sequence is as shown in SEQ ID NO.3. By application of the specific molecular tag to cDNA library construction, the specific molecular tag can be added to a 5' terminal of each cDNA to make each gene unique in a process of reverse transcription of RNA into full-length cDNA, and each gene in samples can be quantified according to sequences of the specific molecular tag in subsequent information analysis.

The present invention is directed to the full-length cDNA sequence encoding human diaphanous-3 (DIAPH3), to DIAPH3 encoded thereby, and to fragments of DIAPH3 and the cDNA. The present invention also provides for the use of the cDNA, and of DIAPH3, as a marker of poor prognosis of breast cancer. Because DIAPH3 appears essential for proper spindle pole formation during mitosis, DIAPH3 is a useful target for screening assays designed to identify inhibitors or modulators of DIAPH3 activity, which are useful for the treatment of cancer, particularly breast cancer. Thus, the invention further provides methods of using DIAPH3, or fragments thereof, in assays to identify such compounds.

The invention relates to a process for generating infectious Newcastle disease virus (NDV) entirely from cloned full-length cDNA and to the use of vaccines and diagnostic assays generated with and derived from the process. The process offers the possibility to modify the NDV genome by means of genetic modification and allows for the introduction of mutations, deletions and/or insertions. The process can be used to modify the virulence of NDV, thus generating new attenuated live vaccines with enhanced properties. The process can be used to modify the antigenic make-up of NDV, to allow the generation of live NDV marker vaccines that can be serologically distinguished from NDV field strains.

The invention discloses a method for cultivating a transgenic plant with improved insect resistance by using RNA interference technology and a special DNA fragment thereof. The DNA fragment is a gene fragment shown as a formula I, wherein a forward sequence (SEQ) is any one fragment which at least comprises 19bp in full-length cDNA fragments of an ATP synthase E subunit gene; a reverse sequence (SEQ) and the forward sequence (SEQ) are mutually reversely complementary; X is a spacer sequence between the forward sequence (SEQ) and the reverse sequence (SEQ), and the X and the forward sequence (SEQ) or the X and the reverse sequence (SEQ) are both not complementary mutually; and the full-length cDNA of the ATP synthase E subunit gene is shown as a sequence 1 in a sequence table. The invention also aims to provide application of the ATP synthase E subunit gene serving as an RNA interference target gene in cultivating the transgenic plant with improved insect resistance. Experiments prove that aphids inoculated to wild-type tobacco grow and reproduce normally; and aphids inoculated to transgenic tobacco starts to die on the fourth day, dead aphid bodies is blackened after five days and no live aphids can be seen after seven days. The formula (I) is forward sequence (SEQ)-X- reverse sequence (SEQ).

The present invention relates to a gene and gene product, SGA-1M that is differentially expressed in cancer tissues and cell lines. Suppression Subtractive Hybridization and microarray screening were used to screen for differential expression of SGA-1M in cancer tissues and cell lines. Expression analysis has demonstrated overexpression of SGA-1M in breast cancer tissue and breast cancer derived cell lines, in ovarian cancer, skin cancer, a cancer of the lymphoid system, thyroid cancer, pancreatic cancer, stomach cancer, and lung cancer. The gene is expressed as a 1.95 kb mRNA. The full length cDNA comprises two open reading frames encoding polypeptides of 221 and 75 amino acids, respectively. Monitoring expression levels of SGA-1M is useful for the diagnosis and prognosis of cancer as well as for evaluating the risk of developing certain types of cancers and the risk of metastasis of cancer. Reagents that target SGA-1M are useful for the treatment of cancer.

The invention provides novel human full-length cDNA clones, novel polynucleotides, related polypeptides, related nucleic acid and polypeptide compositions, and related modulators, such as antibodies and small molecule modulators. The invention also provides methods to make and use these cDNA clones, polynucleotides, polypeptides, related compositions, and modulators. These methods include diagnostic, prophylactic and therapeutic applications. The compositions and methods of the invention are useful in treating proliferative disorders, e.g., cancers, and inflammatory, immune, bacterial, and viral disorders.

The invention discloses a preparation method and an application of a molecular marker of a rape male sterility restoring gene. The preparation method comprises the following steps of A, designing degenerate primers according to PPR gene conservative amino acid sites, B, extracting a lamina DNA of a rape cytoplasm male sterility restoring line through a cetyltrimethyl ammonium bromide (CTAB) method and magnifying the lamina DNA through a touchdown polymerase chain reaction (PCR) process to obtain a specific restoring gene candidate segment, C, carrying out recovery, conversion, sequencing and sequence analysis processes on PCR products, D, carrying out 5 ' RACE and 3 ' RACE processes and splicing a sequencing result to obtain a full-length cDNA sequence, and E, designing specific primers in 5 ' UTR and 3 ' UTR zones of the full-length cDNA sequence according to the full-length cDNA sequence to obtain a molecular marker. The invention also discloses an application of a restoring gene molecular marker in hybrid rape parent breeding. The preparation method has the advantages of improvements of a speed of restorer seeding and an efficiency of verification, simple and easy method, good operability, rapid identification speed of restoring genes, great reduction of necessary workload of test cross and progeny planting observation in a restorative verification process, and improvement of an efficiency of hybrid seeding.

The present invention relates to the cloning, recombination and salt tolerance analysis of cotton Na+/H+ reverse transport protein GhNHX1 gene, and belongs to the field of molecular biology and biological technology. Total RNA is extracted from cotton leaf treated with 0.4 mol/L NaCl solution and inversely transcribed into cDNA. Facultative primer is used for conventional PCR to obtain intermediate segment, and whole length cDNA is obtained through fast amplification in 3' and 5' ends. The gene is transformed to salt-sensitive yeast mutant to restore its salt tolerant capacity to some degree; and the gene is further transformed to tobacco to obtain transgenic plant capabl eof growing normally in salt concentration of 200 mmol/L. Therefore, the gene is one important salt tolerant gene and may be used in raising the salt tolerant capacity of plant for plantation in saline land.