Method, self-replicating vector, assay kit and analysis device
A carrier, a specific technology, applied in the direction of biochemical cleaning devices, biochemical equipment and methods, and the determination/inspection of microorganisms
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Embodiment approach
[0062] Hereinafter, embodiments of the present invention will be described in detail.
[0063] A method of obtaining epigenetic information of a cell according to an embodiment uses a self-replicating reporter nucleic acid construct. When the reporter nucleic acid construct self-replicates in the nucleus of the subject cell, it transcribes the modified state of a specific sequence in the genome of the subject cell into its own sequence. That is, the reporter nucleic acid construct contains the same sequence as the specific sequence to be observed. Furthermore, by self-replication, the modified state on a specific sequence of the genome is transcribed (copied) to the same sequence as that in the reporter nucleic acid construct. Reporter nucleic acid constructs Depending on the presence of functional groups transcribed into the reporter nucleic acid construct, a detectable signal occurs. In this case, the presence or absence or magnitude or amount of the signal differs dependi...
no. 1 Embodiment approach
[0077] In the method for obtaining epigenetic information of cells, a self-replicating vector is used under the condition that a replication initiation protein is expressed in the cell. Self-replicating vectors are also referred to as "reporter vectors".
[0078] use figure 1 , an example of a self-replicating vector will be described. Self-replicating vector 1 contains reporter gene expression unit 2 and replication initiation sequence 3 . The replication initiation sequence 3 and the reporter gene expression unit 2 are present on the same nucleic acid.
[0079] The reporter gene expression unit 2 includes a target nucleic acid sequence 4 , a reporter gene 5 encoding a reporter protein and a transcription termination signal sequence 6 .
[0080] The target nucleic acid sequence 4 is a nucleic acid sequence from which epigenetic information is to be obtained. The target nucleic acid sequence 4 has the same sequence as the specific nucleic acid sequence of the test cell. F...
no. 2 Embodiment approach
[0143] Next, refer to image 3 One example of a plasmid vector will be described.
[0144] Self-replicating vectors31 as plasmid vectors, such as image 3 As shown, it has the target nucleic acid sequence 4, the reporter gene 5 functionally connected downstream thereof, the IRES32 functionally connected downstream thereof, the sequence 33 encoding the replication initiation protein functionally connected downstream thereof, and the functional Transcription termination signal sequence 34 linked to it and replication initiation sequence 3 functionally linked downstream thereof. In this example, the constituent elements of the reporter gene expression unit 2 are the target nucleic acid sequence 4 , the reporter gene 5 and the transcription termination signal sequence 34 . In addition, the constituent elements of the replication initiation protein unit are target nucleic acid sequence 4, IRES32, sequence 33 encoding a replication initiation protein, and transcription termination...
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