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72 results about "Mirna target" patented technology

Novel multifunctional dual-luciferase reporter gene plasmid

The invention discloses a novel multifunctional dual-luciferase reporter gene plasmid. The plasmid contains a luc firefly luciferase gene, a Rluc renilla luciferase gene, a SV40 poly(A) termination signal part and two independent multiple cloning sites, wherein the two independent multiple cloning sites are located on the upstream and the downstream of a luc firefly luciferase gene coder frame respectively. The dual-luciferase reporter gene plasmid containing detection gene and reference gene is successfully constructed, not only can the detection gene be transferred into a subject cell conveniently, but also the reference gene can be taken in correspondingly, the complexity of experiment operation and experiment errors are greatly reduced, the plasmid has the multiple cloning sites at thetwo ends of the firefly luciferase gene, the multiple cloning site located on the upstream of firefly luciferase can introduce a promoter sequence which can be used for genetic transcription adjustment, control and detection, and the multiple cloning site located on the downstream of the firefly luciferase can introduce a 3'UTR sequence which can be used for miRNA target identification, so that switching of different application of the same plasmid is achieved.
Owner:SUN YAT SEN UNIV

Reference internal type dual-luciferase reporter vector and application thereof

The invention discloses a reference internal type dual-luciferase reporter vector and an application thereof. Firefly luciferase gene is fused between the renilla luciferase gene of the pRL-TK vector and the ampicillin resistance gene, the two luciferases are on the same vector, and the firefly luciferase gene is used as reference. The one-step binary bridging coupling long-distance polymerase chain reaction (PCR) and the Escherichia coli vivo homologous recombination method are utilized to place the firefly luciferase gene and the renilla luciferase gene on the same vector; and monoclonal sites are introduced in the 3' untranslated region of the renilla luciferase gene for conveniently cloning the target segment, thus the reference internal type dual-luciferase reporter vector can be constructed. The vector of the invention has the advantages of high repeatability, little multiple-pore variation, convenient operation and precise quantification. The vector of the invention is suitablefor the screening, identification and confirmation of the miRNA target molecule and also suitable for the quantitative analysis of the activity change of miRNA in cellular level.
Owner:INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI

Method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability

The invention discloses a method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability, and relates to the bioengineering technology. The method comprises the following steps: (1) sample collection; (2) sequencing of small RNA; (3) pretreatment of sequencing raw data; (4) sequence alignment; (5) new miRNA prediction; (6) analysis of miRNA differential expression; (7) prediction of differential miRNA target genes; (8) synthesis of miRNA mimic, miRNA inhibitor and control chain; (9) microinjection and artificial feeding of miRNA mimic and miRNA inhibitor to nilaparvata lugens and verification of the function of miRNAs; (10) preliminary study on oryza sativa resistance adaptability change of the nilaparvata lugens after microinjection and artificialfeeding. Small RNAs of two nilaparvata lugens groups are deeply sequenced with a high-throughput sequencing technology, and are analyzed, identified and predicted according to subsequent bioinformatics, differences between the two nilaparvata lugens groups on insect-susceptible oryza sativa TN1 and insect-resistant oryza sativa YHY 15 are compared, the miRNA associated with host resistance adaptability is analyzed, screened and discovered, and the new method is provided for oryza sativa insect-resistant breeding.
Owner:INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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