CN105331621A - Elovl 6 genes of Eriocheir sinensis and clone method of Elovl 6 genes - Google Patents
Elovl 6 genes of Eriocheir sinensis and clone method of Elovl 6 genes Download PDFInfo
- Publication number
- CN105331621A CN105331621A CN201510790826.7A CN201510790826A CN105331621A CN 105331621 A CN105331621 A CN 105331621A CN 201510790826 A CN201510790826 A CN 201510790826A CN 105331621 A CN105331621 A CN 105331621A
- Authority
- CN
- China
- Prior art keywords
- elovl6
- cdna
- mitten crab
- fragment
- gene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to Elovl 6 genes of Eriocheir sinensis and a clone method of the Elovl 6 genes. The method comprises steps as follows: (1), Elovl 6 amino acid sequences of different species are compared, primers Elovl 6-F1/Elovl 6-R1 are designed according to corresponding conservative areas, extracted total RNA (ribose nucleic acid) of the Eriocheir sinensis is reversely transcribed into the first chain of cDNA (complementary deoxyribonucleic acid), a PCR (polymerase chain reaction) is performed with cDNA as a template, and an obtained core fragment is cloned to a carrier and sequenced; (2), a 3'-Elovl6 RACE upstream primer and a 5'-Elovl6 RACE downstream primer are designed respectively according to the core fragment, the extracted total RNA of the Eriocheir sinensis is reversely transcribed into the first chain of 3'-cDNA and the first chain of 5'- cDNA, the 3'-cDNA and the 5'- cDNA are taken as templates, RACE (rapid-amplification of cDNA ends) is performed on a 3' end lateral wing fragment and a 5' end lateral wing fragment, and the obtained 3' end lateral wing fragment and the 5' end lateral wing fragment are connected with the carrier respectively, cloned and sequenced; (3), the obtained end lateral wing fragments and the obtained core fragment are spliced, and the full-length sequence of the Elovl 6 genes of the Eriocheir sinensis is obtained.
Description
Technical field
The present invention relates to genetically engineered field, specifically, relate to mitten crab (Eriocheirsinensis) Elovl6 gene and cloning process thereof.
Background technology
Polyunsaturated fatty acid (Polyunsaturatedfattyacids, PUFAs) refer to containing two or more double bonds and carbon chain lengths more than the lipid acid of 18 carbon atoms, generated through a series of extending enzyme and desaturase catalyzed reaction by lipid acid.PUFAs is the precursor of body biologically active substance, form the main moiety of the material such as organism membrane phospholipid and serum triglyceride, usually in the processes such as aquatic animal to grow, breeding, metabolism, raising food utilization efficiency, play important physiological function.Longer chain fatty acid carbochain extending enzyme (Elongationoflong-chainfattyacids, Elovls) be one of biosynthetic key enzyme of polyunsaturated fatty acid, in catalyze fatty acid extension, play first speed limit effect, longer chain fatty acid (>=C16) and over-long chain fatty acid (>=C20) can be catalyzed and synthesized.
Elovl6 is one of important member in longer chain fatty acid carbochain extending enzyme family, is the microsomal enzyme be positioned in endoplasmic reticulum, usually plays a significant role in lipidic biomass synthesis, fatty acid metabolism and some diseases metabolic process.Elovl6 can extend saturated and monounsaturated fatty acids, and the process of synthesizing stearic acid (C18:0) at catalysis palmitinic acid (C16:0) plays vital effect, for important foundation is established in the generation of PUFA.
Mitten crab (Eriocheirsinensis) is the important economic freshwater aquiculture crab class of China, and belong to Crustachia (Crustacea), Decapoda (Decapoda), occupies critical positions in China's culture fishery.Lipid has a very important role to the growth of mitten crab and the growth of sexual gland, can as the composition of the metabolism energy and necessary tissue and cytolemma.Simultaneously the n-3HUFA such as DHA, EPA is that crab class is grown required important lipid acid, plays important regulating and controlling effect to growth, reproduction, metabolism, immunity etc.Therefore, by obtaining longer chain fatty acid extending enzyme (Elovl) 6 gene, and study its function and be conducive to probing into a dark step of fatty acid synthesis pathway, also do for himself fatty acid synthesis ability power of mitten crab simultaneously and well supplemented, for related work is below laid a good foundation.
Summary of the invention
The object of the present invention is to provide a kind of mitten crab Elovl6 gene.
Another object of the present invention is to provide a kind of albumen of mitten crab Elovl6 genes encoding.
Another object of the present invention is to provide a kind of cloning process of mitten crab Elovl6 gene.
Above-mentioned purpose of the present invention is realized by following technological method:
A kind of mitten crab Elovl6 gene, its nucleotide sequence is as shown in SEQIDNO:1.
A protein for mitten crab Elovl6 genes encoding, its aminoacid sequence is as shown in SEQIDNO:2.
The cloning process of mitten crab Elovl6 gene of the present invention comprises the steps:
(1) the Elovl6 aminoacid sequence of different plant species is compared, according to the conservative region design primer Elovl6-F1/Elovl6-R1 of correspondence, the mitten crab total serum IgE reverse transcription of extraction is become cDNA first chain, as template, carry out PCR reaction, obtain mitten crab Elovl6 gene core fragment, core fragment to be cloned on carrier and to check order;
The nucleotide sequence of Elovl6-F1 is as shown in SEQIDNO:3, and the nucleotide sequence of Elovl6-R1 is as shown in SEQIDNO:4;
(2) according to above-mentioned gained Elovl6 gene core fragment, design 3 '-Elovl6RACE downstream primer and 5 '-Elovl6RACE upstream primer respectively, the mitten crab total serum IgE reverse transcription of extraction is become 3 '-cDNA and 5 '-cDNA first chain, as template, carry out 3 ' and 5 ' end flanking fragment RACE, be connected obtain 3 ' with carrier respectively with 5 ' end flanking fragment, clone and check order;
The nucleotide sequence of 3 '-Elovl6RACE upstream primer is as shown in SEQIDNO:5, and the nucleotide sequence of 5 '-Elovl6RACE downstream primer is as shown in SEQIDNO:6;
(3) by obtain 3 ' and 5 ' end flanking fragment splice with core fragment respectively, obtain the full length sequence of mitten crab Elovl6 extending enzyme gene.
In a preferred embodiment of the invention, step (2) and (3) middle total serum IgE are extracted by Trizol method and obtain from mitten crab hepatopancreas.
In a preferred embodiment of the invention, carrier described in step (2) and (3) is pMD-19T plasmid.
The present invention also provides a kind of carrier, and it comprises mitten crab Elovl6 gene.
The present invention also provides a kind of transfectional cell, and it comprises above-mentioned carrier.
The present invention also provides a kind of transfectional cell, and it expresses above-mentioned albumen.
In the preferred embodiment of the present invention, this transfectional cell is a kind of in yeast saccharomyces cerevisiae, pichia spp, intestinal bacteria, Jin Sirui yeast etc., is preferably yeast saccharomyces cerevisiae.
Compared with existing extending enzyme gene, gene of the present invention has following characteristics:
1) reported first of the present invention separating clone from mitten crab has the gene extending chain fatty acid function, and the albumen of this genes encoding has the activity extending C16:0 and C16:1, plays a role in the upstream phase of longer chain fatty acid building-up process.Especially, make the fall of saturated fatty acid ratio C16:0/C18:0 for maximum in current known Elovl6 gene, this shows that the present invention clones the mitten crab Elovl6 gene obtained is high efficiency gene saturated fatty acid C16:0 being converted into saturated fatty acid C18:0.This is transgenic research exploitation, and such as transgenic microorganism synthesis PUFAs necessary longer chain fatty acid extending enzyme gene, provides a basis.
2) protein of the mitten crab Elovl6 genes encoding of the present invention clone has 290 amino acid, there is typical extending enzyme family feature: the cross-film district of 6 predictions, the redox center Histidine bunch HWYHH of high conservative, multiple conserved regions and ER retention signal KXKXX.The protein molecular weight of prediction is 34.3KD, and theoretical iso-electric point is 9.48.Analyze through Blast in NCBI and show, mitten crab longer chain fatty acid extending enzyme (Elovl) 6 has higher homology with the longer chain fatty acid extending enzyme 6 of other species, such as reach 49%-51% with longer chain fatty acid extending enzyme 6 similarity of people, zebra fish, house mouse, golden head porgy, the highest with the Elovl6 similarity of Environment of Litopenaeus vannamei Low, reach 89%.
The present invention obtains mitten crab Elovl6 full length gene and provides effective platform for furtheing investigate its fatty acid metabolism route of synthesis, have laid a good foundation for optimizing feed formulation, raising mitten crab quality and economic benefit, there is very important theory significance and realistic meaning.
Accompanying drawing explanation
Fig. 1 shows the main process of synthesis of long-chain polyunsaturated fatty acids in organism, wherein Elovl6 extending enzyme is in the upstream of whole pathways metabolism, can by palmitinic acid (C16:0, palmiticacid, PA) stearic acid (C18:0 is extended to, Stearicacid, SA), be transform necessary fatty acid elongase such as generating DHA, EPA.
Fig. 2 is the evolutionary tree of different plant species longer chain fatty acid carbochain extending enzyme 2,5,6.
Fig. 3 is the full length sequence of mitten crab (Eriocheirsinensis) Elovl6 gene.
Fig. 4 is the aminoacid sequence of the protein of mitten crab (Eriocheirsinensis) Elovl6 coded by said gene.
Embodiment
Explain the present invention further below in conjunction with embodiment, but embodiment does not limit in any form to the present invention.
Embodiment 1
Clone mitten crab Elovl6 gene
(1) the Elovl6 aminoacid sequence of pea sprouting, Rattus norvegicus, channel catfish, blue these different plant species of catfish is compared, according to conservative region design primer Elovl6-F1 (its nucleotide sequence is as shown in SEQIDNO:3) and the Elovl6-R1 (its nucleotide sequence is as shown in SEQIDNO:4) of correspondence, Trizol method is utilized to extract the total serum IgE of mitten crab hepatopancreas, be cDNA first chain by it reverse transcription, carry out PCR reaction as template.
PCR application of sample system: cDNA template 1 μ L, dNTPMixture (2.5mmol/L) 4 μ L, each 0.5 μ L, the 10 × PCRbuffer2.5 μ L of above-mentioned two kinds of primers (10 μMs), rTaq enzyme (5U/ μ L) 0.25 μ L, adds aseptic deionized water to cumulative volume 25 μ L.
PCR reaction conditions: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 57 DEG C of annealing 30s, 72 DEG C extend 30s, 32 circulations; 72 DEG C extend 10min, 16 DEG C of preservations.
The mitten crab Elovl6 gene core fragment obtained is cloned into pMD-19T carrier and checks order.
(2) according to above-mentioned gained Elovl6 gene core fragment, design 3 '-Elovl6RACE upstream primer (its nucleotide sequence is as shown in SEQIDNO:5) and 5 '-Elovl6RACE downstream primer (its nucleotide sequence is as shown in SEQIDNO:6), by the mitten crab total serum IgE of extraction respectively reverse transcription become 3 '-cDNA and 5 '-cDNA first chain, as template, carry out 3 ' and 5 ' end flanking fragment RACE:
3 ' terminal fragment rapid amplifying with 3 '-cDNA be template, according to SMARTRACEcDNA amplification kit specification sheets, primer uses UPM (5 '-CTAATACGACTCACTATAGGGC-3 ') and 3 '-Elovl6RACE upstream primer.3 '-RACEPCR reacts application of sample system: 3 '-cDNA template 1.25 μ L, the 3 '-Elovl6RACE upstream primer of 10 μMs and each 0.5 μ L of UPM, 10mMdNTPMix0.5 μ L, 50 × Advantage2PolymeraseMix0.5 μ L, 10 × Advantage2PCRbuffer damping fluid 2.5 Μ l, adds sterilizing deionized water to cumulative volume 25.0 μ L.Reaction conditions: 94 DEG C of 30s, 72 DEG C, 3min, 5 circulations; 94 DEG C of 30s, 70 DEG C of 30s, 72 DEG C of 3min, 5 circulations; 94 DEG C of 30s, 68 DEG C of 30s, 72 DEG C of 3min, 28 circulations;
5 ' end rapid amplifying is template with 5 '-cDNA, illustrates with reference to SMARTRACEcDNA amplification kit specification sheets, primer use UPM (5 '-CTAATACGACTCACTATAGGGC-3 ') and 5 '-Elovl6RACE downstream primer.5 '-RACEPCR reacts application of sample system: 5 '-cDNA template 1.25 μ L, the 5 '-Elovl6RACE downstream primer of 10 μMs and each 0.5 μ L of UPM, 10mMdNTPMix0.5 μ L, 50 × Advantage2PolymeraseMix0.5 μ L, 10 × Advantage2PCRbuffer damping fluid 2.5 Μ l, adds sterilizing deionized water to cumulative volume 25.0 μ L.Reaction conditions: 94 DEG C of 30s, 71.5 DEG C, 3min, 5 circulations; 94 DEG C of 30s, 70 DEG C of 30s, 71.5 DEG C of 3min, 5 circulations; 94 DEG C of 30s, 68 DEG C of 30s, 71.5 DEG C of 3min, 28 circulations;
Obtain 3 ' is connected with pMD-19T respectively with 5 ' end flanking fragment, clones and checks order.
3) by obtain 3 ' and 5 ' end flanking fragment splice with core fragment respectively, acquisition mitten crab Elovl6cDNA full length sequence.
Show through sequential analysis, this Elovl6cDNA total length 2247bp, comprise 3 ' the URT region of 5 ' the URT region of 235bp, the open reading frame of 873bp and 1139bp, 3 ' end has tailing signal AATAAA and ployA tail simultaneously.
290 amino acid of this cDNA sequence coding have typical extending enzyme family feature: the cross-film district of 6 predictions, the redox center Histidine bunch HWYHH of high conservative, multiple conserved regions and ER retention signal KXKXX.The protein molecular weight of prediction is 34.3KD, and theoretical iso-electric point is 9.48.
Show through BLASTp comparison, the Elovl6 similarity of mitten crab Elovl6 aminoacid sequence and people, zebra fish, house mouse, golden head porgy reaches 49%-51%, wherein the highest with the Elovl6 similarity of Environment of Litopenaeus vannamei Low, reaches 89%.
Embodiment 2
The function of checking mitten crab (Eriocheirsinensis) Elovl6 gene
The consisting of of YPD liquid nutrient medium used in the present embodiment: 2% peptone, 1% yeast extract and 2% glucose.Consisting of of S.c.-U liquid nutrient medium used: without the YNB0.67% of amino acid nitrogenous source, aminoacid mixture I (VITAMIN B4, arginine, halfcystine, leucine, Methionin, Threonine and tryptophane) 0.01%, aminoacid mixture II (aspartic acid, Histidine, Isoleucine, methionine(Met), phenylalanine, proline(Pro), Serine, tyrosine and α-amino-isovaleric acid) 0.005% and carbon source 2%.Carbon source used all needs filtration sterilization.
Clone obtains the open reading frame (ORF) of mitten crab Elovl6 gene, then be connected on saccharomyces cerevisiae expression pYES2, obtain Yeast Plasmid pYES-Elovl6, simultaneously with empty carrier pYES2 in the mode shocked by electricity, be transferred to together in yeast saccharomyces cerevisiae, obtain recombination yeast INVScI-Elovl6 and INVScI-pYES2 respectively.30 DEG C of constant temperature culture are after 3 days, and picking positive transformant is inoculated in YPD liquid nutrient medium, 28 DEG C, 200rpm incubated overnight.In containing the INVScI of recombinant plasmid, add 15mlS.c.-U (containing 2% glucose), 30 DEG C, spend the night and shake.By 4 DEG C, incubated overnight base, 1500g, 5min collected by centrifugation is in new pipe.With inducing culture (semi-lactosi containing the 2%) suspension cell of 1-2ml, be then inoculated in the inducing culture of 50ml, 30 DEG C, 2000rpm, cultivate 48-72h.1500g, 5min, 4 DEG C of centrifugal recovery, pour out supernatant liquor, 500 μ L sterilized water suspension cells.Transfer in new centrifuge tube, at full speed centrifugal 30s, removing supernatant liquor, gas chromatographic analysis fatty acid constituents change after lyophilize.What table 1 represented is the measurement result that in recombination yeast INVScI-pYES2 and INVScI-Elovl6, lipid acid forms.
Lipid acid composition in table 1 recombination yeast INVScI-pYES2 and INVScI-Elovl6
Containing four kinds of fatty acid components in yeast saccharomyces cerevisiae INVScI, palmitinic acid (C16:0) respectively, palm monoenoic acid (C16:1n-7), stearic acid (C18:0) and 18 carbon monoenoic acids (C18:1n-9), analyze its content to show: in the recombination yeast INVSc-Elovl6 importing Elovl6 gene, palmitinic acid (C16:0) and Zoomeric acid (C16:1n-7) content are compared with the decline of recombination yeast INVScI-pYES2, and the content of stearic acid (C18:0) and 18 carbon monoenoic acids (C18:1n-9 and C18:1n-7) increases, this shows that the protein of mitten crab Elovl6 genes encoding has the function the saturated of C16 and monounsaturated fatty acids being extended for the saturated of C18 and monounsaturated fatty acids, this conforms to the result of the Elovl6 reported.The albumen of mitten crab Elovl6 genes encoding is Elovl6.
With reporting that different plant species Elovl6 compares, this research not only adopts construction of expression vector system to verify the function of Elovl6 gene first, for its research provides new thinking, and can in Yeast system stably express, for the research in following protein level is had laid a good foundation.
In addition, due to the expression of Elovl6 gene, the saturated fatty acid of C16 is converted into the saturated fatty acid of C18, thus the saturated fatty acid ratio C16:0/C18:0 in fatty acid component is significantly reduced, the albumen of mitten crab Elovl6 of the present invention genetic expression makes the fall of saturated fatty acid ratio C16:0/C18:0 (see table 1) for maximum in current known Elovl6 gene, and this shows that the present invention clones the mitten crab Elovl6 gene obtained is high efficiency gene saturated fatty acid C16:0 being converted into saturated fatty acid C18:0.This is transgenic research exploitation, and such as transgenic microorganism synthesis PUFAs necessary longer chain fatty acid extending enzyme gene, provides a basis, also for optimizing the feed formulation of mitten crab, improving food utilization efficiency and providing a theoretical basis.
Claims (10)
1. a mitten crab Elovl6 gene, its nucleotide sequence is as shown in SEQIDNO:1.
2. an albumen for mitten crab Elovl6 coded by said gene according to claim 1, its aminoacid sequence is as shown in SEQIDNO:2.
3. the cloning process of mitten crab Elovl6 gene according to claim 1, the method comprises the following steps:
(1) the Elovl6 aminoacid sequence of different plant species is compared, according to the conservative region design primer Elovl6-F1/Elovl6-R1 of correspondence, the mitten crab total serum IgE reverse transcription of extraction is become cDNA first chain, as template, carry out PCR reaction, obtain the core fragment of mitten crab Elovl6 gene, described core fragment to be cloned on carrier and to check order;
The nucleotide sequence of Elovl6-F1 is as shown in SEQIDNO:3, and the nucleotide sequence of Elovl6-R1 is as shown in SEQIDNO:4;
(2) according to above-mentioned gained Elovl6 gene core fragment, design 3 '-Elovl6RACE upstream primer and 5 '-Elovl6RACE downstream primer respectively, the mitten crab total serum IgE reverse transcription of extraction is become 3 '-cDNA and 5 '-cDNA first chain, as template, carry out 3 ' and 5 ' end flanking fragment RACE, be connected obtain 3 ' with carrier respectively with 5 ' end flanking fragment, clone and check order;
The nucleotide sequence of 3 '-Elovl6RACE upstream primer is as shown in SEQIDNO:5, and the nucleotide sequence of 5 '-Elovl6RACE downstream primer is as shown in SEQIDNO:6;
(3) by obtain 3 ' and 5 ' end flanking fragment splice with core fragment respectively, obtain the full length sequence of mitten crab Elovl6 extending enzyme gene.
4. method according to claim 3, wherein step (2) and (3) middle total serum IgE are extracted by Trizol method and obtain from mitten crab hepatopancreas.
5. method according to claim 3, wherein carrier described in step (2) and (3) is pMD-19T plasmid.
6. a carrier, it comprises mitten crab Elovl6 gene according to claim 1.
7. a transfectional cell, it comprises carrier according to claim 6.
8. a transfectional cell, it expresses albumen according to claim 2.
9. the transfectional cell according to claim 7 or 8, wherein said cell is a kind of in yeast saccharomyces cerevisiae, pichia spp, intestinal bacteria or Jin Sirui yeast.
10. transfectional cell according to claim 9, wherein said transfectional cell is yeast saccharomyces cerevisiae.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510790826.7A CN105331621B (en) | 2015-11-17 | 2015-11-17 | Eriocheir sinensis Elovl6 gene and its cloning process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510790826.7A CN105331621B (en) | 2015-11-17 | 2015-11-17 | Eriocheir sinensis Elovl6 gene and its cloning process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105331621A true CN105331621A (en) | 2016-02-17 |
| CN105331621B CN105331621B (en) | 2019-01-29 |
Family
ID=55282387
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510790826.7A Expired - Fee Related CN105331621B (en) | 2015-11-17 | 2015-11-17 | Eriocheir sinensis Elovl6 gene and its cloning process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105331621B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110506676A (en) * | 2019-08-23 | 2019-11-29 | 上海海洋大学 | ELOVL1 gene and its application |
| CN110684775A (en) * | 2019-07-01 | 2020-01-14 | 上海海洋大学 | Eriocheir sinensis NKCC gene and cloning method and expression analysis method thereof |
| CN111378669A (en) * | 2020-01-20 | 2020-07-07 | 上海海洋大学 | Eriocheir sinensis 5-HT2B receptor gene and cloning method thereof |
-
2015
- 2015-11-17 CN CN201510790826.7A patent/CN105331621B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| CHEN K.: "Litopenaeus vannamei elongation of very long chain fatty acids protein 6 mRNA, partial cds", 《GENBANK：KP271446.1》 * |
| 姚琴琴 等: "中华绒螯蟹Δ9脂肪酸去饱和酶基因克隆与原核表达", 《中国水产科学》 * |
| 王瑶 等: "中华绒螯蟹RXR基因全长cDNA克隆及表达分析", 《水产学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110684775A (en) * | 2019-07-01 | 2020-01-14 | 上海海洋大学 | Eriocheir sinensis NKCC gene and cloning method and expression analysis method thereof |
| CN110506676A (en) * | 2019-08-23 | 2019-11-29 | 上海海洋大学 | ELOVL1 gene and its application |
| CN111378669A (en) * | 2020-01-20 | 2020-07-07 | 上海海洋大学 | Eriocheir sinensis 5-HT2B receptor gene and cloning method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105331621B (en) | 2019-01-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Investigating long-chain polyunsaturated fatty acid biosynthesis in teleost fish: Functional characterization of fatty acyl desaturase (Fads2) and Elovl5 elongase in the catadromous species, Japanese eel Anguilla japonica | |
| Tanomman et al. | Characterization of fatty acid delta-6 desaturase gene in Nile tilapia and heterogenous expression in Saccharomyces cerevisiae | |
| Lin et al. | Cloning, tissue distribution and nutritional regulation of a fatty acyl Elovl4-like elongase in mud crab, Scylla paramamosain (Estampador, 1949) | |
| Liu et al. | Functional characterization of a Δ5-like fatty acyl desaturase and its expression during early embryogenesis in the noble scallop Chlamys nobilis Reeve | |
| CN103014053B (en) | Synechocystis efficient double homologous recombinant vector as well as construction method and application thereof | |
| Patel et al. | Emerging prospects of microbial production of omega fatty acids: recent updates | |
| CN105189740B (en) | Engineered microorganisms | |
| CN101124330A (en) | Recombinant production docosahexaenoic acid (DHA) in yeast | |
| Kabeya et al. | Modification of the n-3 HUFA biosynthetic pathway by transgenesis in a marine teleost, nibe croaker | |
| CN105331621A (en) | Elovl 6 genes of Eriocheir sinensis and clone method of Elovl 6 genes | |
| CN105368851B (en) | It is a kind of from ω -3 desaturase of phytophthora parasitica, the carrier containing the desaturase, recombinant microorganism and its application | |
| CN1057122C (en) | A recombinant 9 desaturase and a gene encoding the same | |
| Ma et al. | Cloning and characterization of a delta-6 desaturase encoding gene from Nannochloropsis oculata | |
| Lu et al. | Biomass and nutrient productivities of Tetraselmis chuii under mixotrophic culture conditions with various C: N ratios | |
| Xie et al. | Comparsion of activities of fatty acyl desaturases and elongases among six teleosts with different feeding and ecological habits | |
| Kim et al. | Isolation and functional characterization of polyunsaturated fatty acid elongase (AsELOVL5) gene from black seabream (Acanthopagrus schlegelii) | |
| Huo et al. | Outdoor growth characterization of an unknown microalga screened from contaminated Chlorella culture | |
| JP2011505163A5 (en) | ||
| KR20200132835A (en) | High productivity method for growing algae | |
| Oboh | Investigating the long-chain polyunsaturated fatty acid biosynthesis of the African catfish Clarias gariepinus (Burchell, 1822) | |
| CN102226196B (en) | DNA sequence for coding Myrmecia incisa delta-6 fatty acid elongase and application thereof | |
| Kim et al. | Isolation and functional characterization of a delta 6-desaturase gene from the pike eel (Muraenesox cinereus) | |
| CN102260686B (en) | Method for improving content of total grease, linoleic acid or alpha-linolenic acid in chlorella | |
| CN102373229B (en) | Saccharomyces oleaginosus delta12 and delta15 difunctional fatty acid desaturase gene and cloning method thereof | |
| CN109837290A (en) | The application of ShFAD2 gene family and ShFAD3 gene family in the genetically modified plants of initiative high yield ALA |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190129 Termination date: 20191117 |