Nucleic acid sequences and their use as promoters

A nucleotide sequence and promoter technology, applied in the field of genetic engineering, can solve the problems of few genome fragments and little understanding of functions

Active Publication Date: 2021-08-10
SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Relatively little research has been done on this segment of the genome in the early days, and little is known about its function

Method used

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  • Nucleic acid sequences and their use as promoters
  • Nucleic acid sequences and their use as promoters
  • Nucleic acid sequences and their use as promoters

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0082] 1. Plasmid construction

[0083] 1. Construction of pT2AL-Pn-eGFP plasmid

[0084] Firstly, the unique upstream promoter sequence of P. nyererei and the front 48bp coding region sequence of the lg gene (hereinafter referred to as the Pn sequence) are connected into the pT2AL empty vector plasmid to construct the pT2AL-Pn plasmid; then eGFP The fluorescent protein coding region sequence was ligated into the downstream of the Pn sequence in the pT2AL-Pn plasmid constructed above to obtain the pT2AL-Pn-eGFP plasmid.

[0085] Specific steps are as follows:

[0086] 1.1 Construction of pT2AL-Pn plasmid

[0087] 1.1.1 Through the analysis of the multiple cloning site sequence of the empty vector pT2AL (provided by Professor Li Li’s laboratory of Southwest University), find the appropriate double restriction site Sma I, BamH I (NEB) (present in the pT2AL empty vector multiple clone site, but not in the target fragment Pn, so as not to cut the target fragment).

[0088] 1.1...

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Abstract

The invention relates to the technical field of genetic engineering, in particular to a nucleic acid sequence and its application as a promoter. The present invention provides a new promoter sequence, which is a nucleic acid having a nucleotide sequence as shown in SEQ ID NO: 1; or a nucleic acid that substitutes, deletes or adds one or more nucleotides in this fragment; or with it Complementary or partially complementary sequences. The promoter sequence can drive protein expression in various tissues or cells.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a nucleic acid sequence and its application as a promoter. Background technique [0002] Gene expression refers to the process in which a gene produces biologically active RNA or protein through transcription, translation, etc., and expresses its stored genetic information, which is regulated by multiple levels and different factors. Whether in eukaryotes or prokaryotes, most regulatory events occur at the transcriptional level. Transcription is not only the first step in gene expression, but also a key step in controlling gene activity. Promoter is an important participant in the regulation of transcription level and has been paid much attention for a long time. [0003] Promoters are a DNA sequence that RNA polymerase recognizes and binds to initiate transcription. It contains conserved sequences required for RNA polymerase specific binding and transcription initi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/85C12N15/65C07K14/435A61K38/16
CPCA61K38/00C07K14/43595C07K14/461C12N15/65C12N15/85
Inventor 谷浪屿李陈峥毛晓兵韦宗仿黄友葵何希敏周文君李礼王德寿
Owner SUN YAT SEN UNIV
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