CN109055356A - The preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the application in living body interference - Google Patents
The preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the application in living body interference Download PDFInfo
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Abstract
The present invention relates to the preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the applications in living body interference, belong to molecular biology RNA jamming field.It is by extracting the auspicious carp sexual gland RNA of good fortune, and reverse transcription synthesizes dsRNA at cDNA, using the cDNA of reverse transcription, by the dsRNA of intraperitoneal injection synthesis, to inhibit the expression of the auspicious carp igf3 gene of good fortune.The present invention utilizes double-stranded RNA perturbation technique for the first time, and the interference to target gene is realized on living body carp, can effectively inhibit the expression of target gene, and reach good interference effect.Good means are provided for research of the fish functional gene on living body.
Description
Technical field
The present invention relates to the preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the applications in living body interference, belong to
Molecular biology RNA jamming field.
Background technique
Carp is as the main fish culture kind in China, and the progress on gene function is slower, main cause
It is a lack of the living body RNA interference silent technology required for being suitable for growing animal research, with efficient interference effect.Currently,
Limitation of common tiny RNA (siRNA) perturbation technique due to its transfection efficiency, it is difficult to higher interference is maintained on fish living body
Efficiency, and siRNA requirement is big in animal experiment, higher cost.External source and endogenous double-stranded RNA (double-stranded
RNA dsRNA) induce the suppressed phenomenon of gene expression of homologous sequence to be known as RNA interference in the cell.Currently, dsRNA is interfered
Technology has been widely used in the gene functional research of species such as plant, bacterium, insect and shrimp crab, and obtains a series of grind
Study carefully achievement (Waterhouse et al., 1998；Cogoni et al.,1999；Ngo et al.,1998；Lohman et
al.,1999).Start to use in the gene functional research of zebrafish embryo in recent years (Wargelius et al., 1999；Yin-
Xiong Li et al.,2000).But the use on fish living body has not been reported.
Summary of the invention
The object of the present invention is to provide the preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and answering in living body interference
With so that the research for fish gene function provides condition.
A kind of technical solution of the present invention, preparation method of the auspicious carp igf3 gene dsRNA of good fortune, utilizes the auspicious carp igf3 gene of good fortune
Nucleotide sequence prepares dsRNA, includes the following steps:
(1) the auspicious carp gonadal tissue RNA of good fortune is extracted: the acquisition auspicious carp ovary tissue of good fortune, using white (Tianjin) biotechnology in Yuanping City
The ultrapure RNA rapidly extracting kit specification of Co., Ltd's EASYspin tissue/cell carries out RNA extraction, passes through 1.2% agar
Sugared detected through gel electrophoresis RNA mass measures RNA concentration using ultraviolet specrophotometer, and is stored in -80 DEG C.
(2) preparation of igf3 gene dsRNA: according to the cDNA sequence of the auspicious carp igf3 gene of good fortune designed for preparation dsRNA
Primer it is a pair of: forward primer SEQ ID NO.1 and reverse primer SEQ ID NO.2；
Forward primer SEQ ID NO.1:
5’-TAATACGACTCACTATAGGGTGTACTGCGTCCTGATCCTG-3'；
Reverse primer SEQ ID NO.2:
5’-TAATACGACTCACTATAGGGGAAGGTTGCTGCTGTGTTGA-3'；, wherein underscore part is opened for T7
Promoter sequences.
Purpose piece is obtained by PCR using the cDNA of the auspicious carp ovary total serum IgE reverse transcription synthesis of good fortune as template to primer with this
Section, then Thermo Scientific TranscriptAid T7 High Yield is used by template of this section of segment
Transcription Kit (U.S.) kit synthesizes dsRNA.
The dsRNA that above-mentioned steps synthesize is used for the interference of carp living body, described method includes following steps:
(1) raising of the auspicious carp fry of good fortune: the auspicious carp fry of good fortune (average weight: 10 ± 1.28g) is raised in circulating water cultivation
In system, the specification of round breeding barrel is diameter 90cm, high 80cm, and natural lighting feeds compound feed for carps, feeds 3 daily
Secondary, each feeding volume is the 5% of weight.
(2) Active MnO2 of dsRNA:
A, injection site selects: the dsRNA of synthesis being diluted to required concentration, according to the note of 5 μ g/g (concentration/fish body weight)
It penetrates concentration and carries out injection site selection.Experiment is divided into 4 groups (n=10): tail vein injection group, intraperitoneal injection group, intramuscular injection group
And control group, every group of 3 repetitions, the detection death rate and jamming effectiveness after injection hour for 24 hours, binding operation difficulty, so that it is determined that
Optimal injection position is abdominal cavity.
B, various dose interference test: 3 dsRNA dosage levels of setting study interference rule: low dose group (1 μ g/g, n
=15), middle dose group (5 μ g/g, n=15) and high dose group (15 μ g/g, n=15), every group of 3 repetitions.Each dosage is set
Control group is set, injects the DEPC water of equivalent respectively.Distinguish 1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d, 11d and 13d after injection
Sexual gland is taken, detecting optimum jamming dosage by the extraction of RNA and RT-q PCR is 5 μ g/g.
(3) RNA jamming effectiveness detects: being injected according to above-mentioned injection site the selection result, experiment is divided into two groups of (n=
20).Experimental group is injected by optimal dose, and control group injects the DEPC water of equivalent, distinguishes 1d, 2d, 3d, 4d, 5d after injection,
6d, 7d, 9d, 11d and 13d take the mixing sample (n=3) of sexual gland, determine interference effect by the extraction of RNA and RT-q PCR detection
It is 7 days between seasonable.
Beneficial effects of the present invention: compared with the control group, the destination gene expression amount in experimental group is substantially less than the present invention
Control group reaches as high as 85% or more.The RNA living body interference carried out using this method has efficient jamming effectiveness, Ke Yiyou
The interference of the realization target gene of effect.
Detailed description of the invention
The double-stranded RNA figure of the auspicious carp igf3 gene of Fig. 1 good fortune.
Fig. 2 is used to detect the real time fluorescent quantitative figure of RNA jamming effectiveness.
Fig. 3: the real time fluorescent quantitative figure of optimal injection position jamming effectiveness.
Fig. 4: the real time fluorescent quantitative figure of optimal injection dosage jamming effectiveness.
C is control control group in Fig. 2-4.
Specific embodiment
Below with reference to embodiment, specific embodiments of the present invention will be further explained, but does not constitute to limit of the invention
System.
1 carp igf3 gene dsRNA of embodiment preparation
1, carp ovary tissue RNA is extracted: the acquisition auspicious carp ovary tissue of good fortune, limited using white (Tianjin) biotechnology in Yuanping City
The ultrapure RNA rapidly extracting kit specification of company's EASYspin tissue/cell carries out RNA extraction, solidifying by 1.2% agarose
Gel electrophoresis detects RNA mass, measures RNA concentration using ultraviolet specrophotometer, and be stored in -80 DEG C.
The double-stranded RNA figure of the auspicious carp igf3 gene of good fortune is as shown in Figure 1.
2, dsRNA design of primers: being classified as foundation with carp igf3 gene (GenBank:KT895500.1) nucleotides sequence,
The online primer-design software of dsRNA (http://www.flyrnai.org/cgi-bin/RNAi_ is used in open reading frame
Find_primers.pl) a pair of designed for preparing the primer of double-stranded RNA:
Forward primer SEQ ID NO.1:
5’-TAATACGACTCACTATAGGGTGTACTGCGTCCTGATCCTG-3'；
Reverse primer SEQ ID NO.2:
5’-TAATACGACTCACTATAGGGGAAGGTTGCTGCTGTGTTGA-3'；Wherein underscore part is T7 starting
Subsequence.
3, dsRNA is synthesized:
Passed through using the dsRNA primer of design synthesis using the cDNA of the auspicious carp ovary total serum IgE reverse transcription synthesis of good fortune as template
PCR obtains target fragment, then uses TranscriptAid T7 High Yield by template of this section of segment
Transcription Kit (Thermo Scientific, USA) kit synthesizes dsRNA, specific as shown in table 1:
Table 1
| DEPC-treated water | to 20μL |
| 5X TranscriptAid Reaction Buffer | 4μL |
| ATP/CTP/GTP/UTP mix* | 8μL |
| Template DNA | 2μL |
| TranscriptAid Enzyme Mix | 2μL |
| Total volume | 20μL |
PCR reaction condition is as follows: 37 DEG C of 2h.
4, dsRNA is purified:
The 3M sodium acetate of 115 μ LDEPC water and 15 μ L is added in 20 μ L reaction systems, mixes；Then isometric chlorine is added
Imitative/phenol (1:1) mixture, is extracted twice, and collects upper phase into new centrifuge tube；The anhydrous second of two volumes is added
Alcohol, -20 DEG C of freezing 30min collect precipitating；DsRNA, -80 DEG C of guarantors are dissolved with DEPC water with after 70% and ethanol washing precipitating
It deposits spare.
The Active MnO2 of 5.dsRNA
(1) injection site selects
The dsRNA of synthesis is diluted to required concentration, is injected according to the injection concentration of 5 μ g/g (concentration/fish body weight)
Position selection.Experiment is divided into 4 groups (n=10): tail vein injection group, intraperitoneal injection group, intramuscular injection group and control group, and every group 3
A repetition detects the death rate and jamming effectiveness, binding operation difficulty after injecting hour for 24 hours, so that it is determined that optimal injection position is
Abdominal cavity (Fig. 3).
(2) various dose interference test
3 dsRNA dosage level research interference rules: low dose group (1 μ g/g, n=15), middle dose group (5 μ g/ are set
G, n=15) and high dose group (15 μ g/g, n=15), every group of 3 repetitions.Control group is arranged in each dosage, respectively injection etc.
The DEPC water of amount.1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d, 11d and 13d take sexual gland after injection respectively, pass through the extraction of RNA
And RT-q PCR detection optimum jamming dosage is 5 μ g/g (Fig. 4).
(3) time dependence interference test
It is injected according to above-mentioned injection site the selection result, experiment is divided into two groups (n=20).Experimental group presses optimal dose
Injection, control group inject the DEPC water of equivalent, and 1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d, 11d and 13d take after injection respectively
The mixing sample (n=3) of sexual gland is detected by the extraction of RNA and RT-qPCR and determines that the disturbing effect time is 7 days (Fig. 2).
Embodiment 2: quantitative fluorescent PCR verifies igf3 gene jamming effectiveness
1, tissue RNA is extracted and cDNA is synthesized
Experimental group and control group gonadal tissue are taken respectively, using white (Tianjin) the RNA rapidly extracting kit extractability in Yuanping City
Glandular tissue RNA, and use the PrimeScript of Takara companyTMRT Master Mix (Perfect Real Time) reagent
For box by RNA reverse transcription at cDNA, reverse transcription step is as shown in table 2.
Table 2
| 5×PrimeScript RT Master Mix(Perfect Real Time) | 2μL |
| Total RNA | 500ng |
| RNase Free dH2O | up to 10μL |
PCR reaction condition is as follows: 37 DEG C of 15min, 85 DEG C of 5s.
2, real-time fluorescence quantitative PCR detects
Igf3 gene jamming effectiveness is detected using Real-Time Fluorescent Quantitative PCR Technique, is made of carp β-actin gene
Internal reference, PCR reaction system are as shown in table 3:
Table 3
| SYBR Premix Ex Taq II(Tli RNaseH Plus)(2×) | 12.5μL |
| PCR Forward Primer(10μM) | 1μL |
| PCR Reverse Primer(10μM) | 1μL |
| DNA profiling (< 100ng) | 2μL |
| DH2O (sterile purified water) | 8.5μL |
| Total | 25μL |
PCR reaction condition is as follows: 95 DEG C of 30s；95 DEG C of 5s, 63 DEG C of 30s, 40 circulations.Experimental result uses 2-ΔΔCtIt calculates
Gene relative expression quantity.
The auspicious carp igf3 gene by fluorescence quantitative PCR the primer of good fortune is as shown in table 4:
Table 4
| igf3 F | AGCAGCGATACCAGAAGCAT |
| igf3 R | GAGTTCCACCGGTAAAGCGT |
| β-actin F | GCTATGTGGCTCTTGACTTCGA |
| β-actin R | CCGTCAGGCAGCTCATAGCT |
6, experimental result
Shown in Fig. 1, for the double-stranded RNA figure of synthesis, dsRNA band is clear, high specificity.
Shown in Fig. 2, the influence situation after igf3-dsRNA to igf3 gene expression amount is injected intraperitoneally for external source.
Influence situation shown in Fig. 3, after different injection site injection igf3-dsRNA to igf3 gene expression amount.
Influence situation shown in Fig. 4, after different injection concentration injection igf3-dsRNA to igf3 gene expression amount.
Compared with the control group, the destination gene expression amount in experimental group is substantially less than control group, reaches as high as 85% or more.
The RNA living body interference carried out using this method has efficient jamming effectiveness, can effective experiment purpose gene silencing.
Note: 1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d, 11d, 13d are respectively 1,2,3,4,5 injected after igf3-dsRNA,
6,7,9,11 and 13 days (sample size >=3).
Sequence table
<110>preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the application in living body interference
<120>preparation method of the auspicious carp igf3 gene dsRNA of good fortune a kind of and the application in living body interference
<141> 2018-08-08
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 40
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
taatacgact cactataggg tgtactgcgt cctgatcctg 40
<210> 2
<211> 40
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 2
taatacgact cactataggg gaaggttgct gctgtgttga 40
Claims (4)
1. a kind of preparation method of the auspicious carp igf3 gene dsRNA of good fortune, it is characterized in that steps are as follows:
(1) the auspicious carp gonadal tissue RNA of good fortune is extracted: the acquisition auspicious carp ovary tissue of good fortune extracts ovary RNA by RNA extracts kit,
By 1.2% agarose gel electrophoresis detect RNA mass, using ultraviolet specrophotometer measure RNA concentration, then reverse transcription at
cDNA；
(2) preparation of igf3 gene dsRNA: according to the cDNA sequence of the auspicious carp igf3 gene of good fortune designed for preparing drawing for dsRNA
Object is a pair of: forward primer SEQ ID NO.1 and reverse primer SEQ ID NO.2；It is total with the auspicious carp ovary of good fortune using the primer pair
The cDNA of RNA reverse transcription synthesis is template, obtains target fragment by PCR, then synthesize using this section of segment as template with kit
dsRNA。
2. the preparation method of the auspicious carp igf3 gene dsRNA of good fortune as described in claim 1, it is characterized in that: the primer pair are as follows:
Forward primer SEQ ID NO.1:
5’-TAATACGACTCACTATAGGGTGTACTGCGTCCTGATCCTG-3'；
Reverse primer SEQ ID NO.2:
5’-TAATACGACTCACTATAGGGGAAGGTTGCTGCTGTGTTGA-3'；
Wherein underscore part is T7 promoter sequence.
3. dsRNA described in claim 1 is for the auspicious carp living body igf3 gene RNA interference of good fortune, it is characterized in that steps are as follows:
(1) raising of the auspicious carp fry of good fortune: by the auspicious carp fry raising of good fortune in circulating water culture system, natural lighting feeds carp
Mixed feed is fed 3 times daily, and each feeding volume is the 4%-6% of weight；
(2) Active MnO2 of dsRNA: intraperitoneal injection is interfered with the concentration that concentration/fish body restatement is 5 μ g/g, control group
Inject same amount of normal saline.
4. dsRNA as claimed in claim 3 is for the auspicious carp living body igf3 gene RNA interference of good fortune, it is characterized in that RNA jamming effectiveness is examined
Survey method is as follows: 1d, 2d, 3d, 4d, 5d, 6d, 7d, 9d, 11d and 13d after dsRNA intraperitoneal injection acquire sexual gland respectively
Tissue detects igf3 gene expression dose using real time fluorescent quantitative method, detects jamming effectiveness, determine that interference time can hold
It is 7 days continuous.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110157743A (en) * | 2019-05-28 | 2019-08-23 | 中国水产科学研究院黄海水产研究所 | For striking the injection and application method of low turbot 14-3-3 gene expression |
| CN111217902A (en) * | 2020-01-20 | 2020-06-02 | 中国水产科学研究院淡水渔业研究中心 | Preparation and application of carp insulin-like growth factor IGF3 recombinant protein |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105296499A (en) * | 2015-11-03 | 2016-02-03 | 中国水产科学研究院淡水渔业研究中心 | Cyrinus carpio IGF3 gene as well as amplification primer group and amplification method thereof |
-
2018
- 2018-08-08 CN CN201810896720.9A patent/CN109055356A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105296499A (en) * | 2015-11-03 | 2016-02-03 | 中国水产科学研究院淡水渔业研究中心 | Cyrinus carpio IGF3 gene as well as amplification primer group and amplification method thereof |
Non-Patent Citations (3)
| Title |
|---|
| FEIBIAO SONG等: "A Novel igf3 Gene in Common Carp (Cyprinus carpio): Evidence for Its Role in Regulating Gonadal Development", 《PLOS ONE》 * |
| 李法君: "青虾IAG及相关基因的克隆及调控关系的研究", 《中国博士学位论文全文数据库农业科技辑》 * |
| 聂永佳: "《渔用药物及鱼病防治手册》", 30 June 1998, 湖南科学技术出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110157743A (en) * | 2019-05-28 | 2019-08-23 | 中国水产科学研究院黄海水产研究所 | For striking the injection and application method of low turbot 14-3-3 gene expression |
| CN111217902A (en) * | 2020-01-20 | 2020-06-02 | 中国水产科学研究院淡水渔业研究中心 | Preparation and application of carp insulin-like growth factor IGF3 recombinant protein |
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