CN113897324A - JcVIPP1 recombinant escherichia coli used as anti-manganese agent and construction method thereof - Google Patents
JcVIPP1 recombinant escherichia coli used as anti-manganese agent and construction method thereof Download PDFInfo
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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Abstract
The invention provides a JcVIPP1 recombinant escherichia coli used as an anti-manganese agent and a construction method thereof, wherein the JcVIPP1 recombinant escherichia coli is constructed by recombining a plastid 1 vesicle inducible protein gene JcVIPP1 in jatropha curcas into a vector and introducing the recombinant escherichia coli. The engineering bacterium of Escherichia coli constructed by the invention has wide application range, can quickly grow after being delayed at normal temperature, has the optimal growth temperature of 37 ℃, and ensures that the bacterium can process Mn at high environmental temperature2+The eutrophication pollution of the water body is removed while the pollution is caused, and the cost is very low; the growth speed of the thalli is high, so that the pollution removal period is short; the produced thallus is further concentrated, and the absorbed Mn can be easily extracted and purifiedHas the advantages of biological smelting.
Description
Technical Field
The invention belongs to the technical field of biochemistry, and particularly relates to JcVIPP1 recombinant escherichia coli used as an anti-manganese agent and a construction method thereof.
Background
VIPP1 was first discovered in peas in 1994 and was named membrane-associated protein (IM 30) due to its binding to chloroplast envelope (envelope) and Thylakoid (THYLAKOID) membranes and a molecular weight of 30kDa, and was also known as IM30(30kDa inner-membrane association protein). In 2001, it was renamed as a Vesicle-inducing Protein in plastid 1 (Vesicle-inducing Protein in plastics 1, VIPP 1). Phylogenetic analyses showed that VIPP1 originated from prokaryotes and possibly evolved from bacteriophage Shock Protein A (PspA). This is the first plant homologous protein of E.coli (Escherichia coli). VIPP1 is involved in the formation or remodeling of green plants and blue algae thylakoid membranes in a manner that changes local structures. In plants containing chloroplasts, VIPP exerts its function by relying on magnesium ions. The experiment of expressing VIPP-his in the in vitro experiment of Escherichia coli shows that VIPP1 depends on combination with GTP to play the role of GTP enzyme. Thereby driving the motion of VIPP1 and maintaining the transmembrane proton gradient of the escherichia coli cell membrane. This may have some relevance to fusion repair of plasma membranes. Meanwhile, experimental research proves that VIPP1 can rescue defective proton leakage in the Escherichia coli PspA mutant and play a role in membrane repair. Under the condition of membrane damage in adversity stress, VIPP1 participates in bacterial stress reaction, specifically interacts with the membrane surface with negative charges, triggers membrane fusion, and achieves the effect of membrane stabilization.
Manganese is a trace element essential to living beings. It is an important component of superoxide dismutase, phosphopyruvate decarboxylase, arginase, glutaminase, etc., and manganese plays an important role in electron transfer in water splitting of photosynthesis. Because of the important role of manganese, researchers have historically focused on the effects of Mn deficiency. Biological manganese deficiency can cause various adverse symptoms, but with the development of industry and agriculture, the use of Mn is increasingly common. In industry, Mn is one of important alloying elements, and is incorporated into various steel materials to increase the hardness of the materials, and thus, both welders and smelters are exposed to the harsh environment. Manganese from manganese mines permeates into soil and water to cause Mn pollution in the environment due to mining, storage and transportation. In agriculture, Mn-containing pesticides such as maneb and mancozeb are used for crop antibiosis, enter the environment and crop bodies through various application modes, and are taken by people. In addition, with the rapid development of economy and traffic, organolead, which has been used as an antiknock agent in the past, is now substituted with MMT (methylcyclopentadienyl manganese tricarbonyl), and can increase the octane number of oil. Thus, Mn enters the engine body through the contact of automobile exhaust and oil.
The harm of excessive manganese to organisms is most reported at present to be human. Mn can enter the human body through 3 pathways: food enters a human body through the digestive tract: secondly, the blood-brain barrier is penetrated into the brain through a respiratory tract; thirdly, the manganese-containing substance enters the human body through skin contact. The toxicity of Mn is mainly to damage globus pallidus and substantia nigra of basal ganglia, influence the secretion and degradation of dopamine, and produce symptoms like ataxia of Parkinson's syndrome and the like. Because of the similarity to iron, excess manganese will also compete for Fe2+The binding site of (a) causes iron deficiency anemia. In addition, excessive Mn can also cause hearing loss, stillbirth, cardiopulmonary insufficiency, syndactyly, and the like.
At present, the existing water body heavy metal pollution treatment methods include adsorption, ion exchange, precipitation/coprecipitation, various separation technologies (filtration, membrane, even magnetic methods), biological methods and natural remediation. Wherein the ion exchange technique is via H carried on a resin+The ion exchange capacity needs to be recovered periodically after a certain time, so that the cost is high; chemical precipitation results in the formation of a large amount of sludge, which is costly and difficult to handle. Membrane processing techniques and electrochemical methods are costly. Phytoremediation, limited by environmental conditions, takes too long。
Disclosure of Invention
The invention aims to provide JcVIPP1 recombinant escherichia coli used as an anti-manganese agent and a construction method thereof, and the constructed escherichia coli engineering bacteria have wide application range, can quickly grow after being delayed at normal temperature, have the optimal growth temperature of 37 ℃, ensure that bacteria can process Mn at high environmental temperature2+The eutrophication pollution of the water body is removed while the pollution is caused, and the cost is very low; the growth speed of the thalli is high, so that the pollution removal period is short; after the generated thalli are further concentrated, the absorbed Mn can be easily refined, and the method has the advantages of biological smelting.
The invention provides a JcVIPP1 recombinant escherichia coli used as an anti-manganese agent, which is constructed by recombining a plastid 1 vesicle-induced protein gene JcVIPP1 in jatropha curcas into a vector and introducing the recombinant escherichia coli.
The invention also provides a construction method of the JcVIPP1 recombinant escherichia coli, which comprises the following steps:
with primer Vipp-F5'-CCG ATGGCCGCAAAATCACAGTTAATT AC-3'; performing RT-PCR on the drought-treated Jatropha curcas cDNA library to obtain a JcVIPP1 full-length coding gene by Vipp-R5'-CCG agcaatcagaattcgTTAGAATTCCTTTCTCTTTTGTCTTAATTCATTG-3', wherein the coding gene has a full length of 1005bp and comprises the following components:
ATGGCCGCAAAATCACAGTTAATTACAGGATTGACCTTGCCATTGCCACCGCCGCATTCTTCCACTTCCTCAACCTCCAATAACAGCAGCAACACTCTCTGTATGGTCAAGCGGCCGCAACTTACGACTTCGTTTTTCAATGGCGGAGTTGAAGCTCTAAAATTTTCTAGGATAAGGACTTGTTCTACTAGGTCCCATTGCTACAGACAAGGTGGAGGTGCTCTTGGCACTCGTATGAATCTTTTTGATCGGTTTGCTAGAGTTGTCAAGTCATATGCAAATGCAATCTTGAGTGGTTTTGAGGACCCGGAAAAAATTCTAGATCAGACGGTTCTTGAAATGAATGATGACTTGACAAAGATGCGTCAGGCCACAGCACAAGTATTGGCATCTCAAAAACGTTTGGAAAATAAATACAAAGCTGCGGAACAAGCTTCTGAGGATTGGTACCGTAAAGCACAACTTGCTCTTCAGAAAGGAGAGGAAGATCTCGCTCGGGAAGCTCTTAAGAGGCGTAAATCTTATGCTGACAATGCAAATTCCTTGAGAGCTCAACTTGATCAACAGAAAAGTGTTGTTGAGAATCTTGTCTCTAATACTCGGCTTTTGGAGAGCAAGATACAGGAGGCAAAGTCTAAAAAAGATACTCTGAAAGCGCGTGCCCAATCTGCAAAAACTCAAACCAAAGTGAATGAGATGCTGGGGAATGTAAATACAAGCAACGCTCTTTCAGCTTTCGAGAAAATGGAAGAGAAAGTATTGCAAATGGAATCAGAAGCTGAAGCACTTGGCCAGTTAGCTACAAGTGAATTGGATGGAAAGTTTGCTTTACTTGAGAGCTCATCTGTTGATGATGATCTTGAGAACCTGAAGAAGGAAATTTCTGGTAGCAAAAAGAGAGGAGAACTGCCGCCCGGTAGAACAGTTGTCAGCAGCTCTGCATTGAGAGATCCTGAGATTGAGATGCAGCTCAATGAATTAAGACAAAAGAGAAAGGAATTCTAA；
encodes a protein consisting of 334 amino acid residues, the amino acid composition is shown in Table 1:
TABLE 1 amino acid composition of JcVIPP1
| Amino acids | Number of | Percentage of | Amino acids | Number of | Percentage of |
| Ala(A) | 31 | 9.30％ | Leu(L) | 39 | 11.70％ |
| Arg(R) | 22 | 6.60％ | Lys(K) | 30 | 9.00％ |
| Asn(N) | 18 | 5.40％ | Met(M) | 9 | 2.70％ |
| Asp(D) | 15 | 4.50％ | Phe(F) | 9 | 2.70％ |
| Cys(C) | 3 | 0.90％ | Pro(P) | 9 | 2.70％ |
| Gln(Q) | 20 | 6.00％ | Ser(S) | 35 | 10.50％ |
| Glu(E) | 29 | 8.70％ | Thr(T) | 20 | 6.00％ |
| Gly(G) | 15 | 4.50％ | Trp(W) | 1 | 0.30％ |
| His(H) | 2 | 0.60％ | Tyr(Y) | 5 | 1.50％ |
| Ile(I) | 7 | 2.10％ | Val(V) | 15 | 4.50％ |
The product obtained by the amplification of the primers is cut by restriction enzymes Hind III and BamH I, then is connected to a vector pYES2, and is introduced into Escherichia coli DH5 alpha to construct JcVIPP1 recombinant Escherichia coli.
Compared with the prior art, the invention has the beneficial effects that:
mn in a clean environment2+In the aspect of pollution, the invention has the advantages of low use cost, wide application and high speed. Compared with the ion exchange technology in the prior art, the ion exchange technology needs to recover the ion exchange capacity periodically, so that the investment of time, manpower, material resources and financial resources is needed, and the engineering bacteria constructed by using the JcVIPP1 has low use cost. Chemical precipitation methods form a lot of sludge, and are also costly and difficult to handle. Membrane processing techniques and electrochemical methods are costly. Phytoremediation, limited by environmental conditions, takes too long.
The engineering bacterium of Escherichia coli constructed by the invention has an adaptive rangeWide range, can quickly grow after being delayed at normal temperature, has the optimal growth temperature of 37 ℃, and ensures that the thalli can process Mn at high temperature in the environment2+The eutrophication pollution of the water body is removed while the pollution is caused, and the cost is very low; the growth speed of the thalli is high, so that the pollution removal period is short; after the generated thalli are further concentrated, the absorbed Mn can be easily refined, and the method has the advantages of biological smelting.
Drawings
FIG. 1 shows Mn of strains obtained by screening in an example of the present invention2+A comparison graph of the minimum inhibitory concentration experiment of (c);
FIG. 2 is a comparative graph of an experiment in one example of the present invention under the condition of a normal LB medium culture;
FIG. 3 shows Mn at 25mM in one embodiment of the invention2+Experimental comparison graphs under the condition of medium culture;
FIG. 4 shows the effect of CAT activity of JcVIPP1 E.coli under manganese stress in accordance with one embodiment of the present invention;
FIG. 5 shows the effect of JcVIPP1 on E.coli POD activity in one embodiment of the present invention;
FIG. 6 shows the effect of JcVIPP1 on the SOD activity of E.coli in one embodiment of the present invention.
Detailed Description
The present invention is described in detail with reference to the embodiments shown in the drawings, but it should be understood that these embodiments are not intended to limit the present invention, and those skilled in the art should understand that functional, methodological, or structural equivalents or substitutions made by these embodiments are within the scope of the present invention.
This example provides a recombinant E.coli of JcVIPP1 used as an anti-manganese agent, in which the gene JcVIPP1 of the vesicle-inducing protein (vesicular-inducing protein in plastics 1, VIPP1) in Jatropha curcas (Jatropha curcas) was recombined into a vector and introduced into E.coli (the strain name is abbreviated as JcV), having resistance to high concentration of manganese ions (Mn)2+) The characteristics of (1) are that Peroxidase (POD), Catalase (Catalase, CAT) and superoxide are contained in an oxidation resisting system in thalli through a growth curveChanges in Superoxide Dismutase (SOD); in contrast to the control, the JcVIPP1 recombinant e.coli has an effect on environmental repair under manganese stress.
The engineering bacterium of Escherichia coli constructed by the invention has wide application range, can quickly grow after being delayed at normal temperature, has the optimal growth temperature of 37 ℃, and ensures that the bacterium can process Mn at high environmental temperature2+The eutrophication pollution of the water body is removed while the pollution is caused, and the cost is very low; the growth speed of the thalli is high, so that the pollution removal period is short; after the generated thalli are further concentrated, the absorbed Mn can be easily refined, and the method has the advantages of biological smelting.
The construction method of the JcVIPP1 recombinant escherichia coli comprises the following steps: with primer Vipp-F5'-CCG tgatacatatgcccg ATGGCCGCAAAATCACAGTTAATTAC-3'; and performing RT-PCR on the drought-treated Jatropha curcas cDNA library to obtain a JcVIPP1(JcMAP30) full-length coding gene by Vipp-R5'-CCG agcaatcagaattcg TTAGAATTCCTTTCTCTTTTGTCTTAATTCATTG-3'. The total length of the coding gene is 1005bp (as follows), and the coding gene codes protein consisting of 334 amino acid residues, and the amino acid composition is shown in table 1. The product amplified from the above primers was digested with restriction enzymes Hind III and BamH I, ligated to vector pYES2(ThermoFisher, USA), and introduced into E.coli DH5 α.
The strains containing the recombinant plasmid were selected with a selection medium. Firstly, screening positive clones by LB solid culture medium containing 50mg/l ampicillin (Amp), carrying out colony PCR by using Vipp-F and Vipp-R primers, and verifying whether the positive clones are false positive or not after agarose gel electrophoresis of PCR products. After the positive colony is subjected to secondary single colony streaking culture and purification, the positive colony is subjected to Mn-containing2+And (4) performing secondary screening on the LB solid culture medium, performing PCR verification on the positive clone again, sequencing a PCR product, and confirming that the gene sequence is correct.
ATGGCCGCAAAATCACAGTTAATTACAGGATTGACCTTGCCATTGCCACCGCCGCATTCTTCCACTTCCTCAACCTCCAATAACAGCAGCAACACTCTCTGTATGGTCAAGCGGCCGCAACTTACGACTTCGTTTTTCAATGGCGGAGTTGAAGCTCTAAAATTTTCTAGGATAAGGACTTGTTCTACTAGGTCCCATTGCTACAGACAAGGTGGAGGTGCTCTTGGCACTCGTATGAATCTTTTTGATCGGTTTGCTAGAGTTGTCAAGTCATATGCAAATGCAATCTTGAGTGGTTTTGAGGACCCGGAAAAAATTCTAGATCAGACGGTTCTTGAAATGAATGATGACTTGACAAAGATGCGTCAGGCCACAGCACAAGTATTGGCATCTCAAAAACGTTTGGAAAATAAATACAAAGCTGCGGAACAAGCTTCTGAGGATTGGTACCGTAAAGCACAACTTGCTCTTCAGAAAGGAGAGGAAGATCTCGCTCGGGAAGCTCTTAAGAGGCGTAAATCTTATGCTGACAATGCAAATTCCTTGAGAGCTCAACTTGATCAACAGAAAAGTGTTGTTGAGAATCTTGTCTCTAATACTCGGCTTTTGGAGAGCAAGATACAGGAGGCAAAGTCTAAAAAAGATACTCTGAAAGCGCGTGCCCAATCTGCAAAAACTCAAACCAAAGTGAATGAGATGCTGGGGAATGTAAATACAAGCAACGCTCTTTCAGCTTTCGAGAAAATGGAAGAGAAAGTATTGCAAATGGAATCAGAAGCTGAAGCACTTGGCCAGTTAGCTACAAGTGAATTGGATGGAAAGTTTGCTTTACTTGAGAGCTCATCTGTTGATGATGATCTTGAGAACCTGAAGAAGGAAATTTCTGGTAGCAAAAAGAGAGGAGAACTGCCGCCCGGTAGAACAGTTGTCAGCAGCTCTGCATTGAGAGATCCTGAGATTGAGATGCAGCTCAATGAATTAAGACAAAAGAGAAAGGAATTCTAA。
TABLE 1 amino acid composition of JcVIPP1
| Amino acids | Number of | Percentage of | Amino acids | Number of | Percentage of |
| Ala(A) | 31 | 9.30％ | Leu(L) | 39 | 11.70％ |
| Arg(R) | 22 | 6.60％ | Lys(K) | 30 | 9.00％ |
| Asn(N) | 18 | 5.40％ | Met(M) | 9 | 2.70％ |
| Asp(D) | 15 | 4.50％ | Phe(F) | 9 | 2.70％ |
| Cys(C) | 3 | 0.90％ | Pro(P) | 9 | 2.70％ |
| Gln(Q) | 20 | 6.00％ | Ser(S) | 35 | 10.50％ |
| Glu(E) | 29 | 8.70％ | Thr(T) | 20 | 6.00％ |
| Gly(G) | 15 | 4.50％ | Trp(W) | 1 | 0.30％ |
| His(H) | 2 | 0.60％ | Tyr(Y) | 5 | 1.50％ |
| Ile(I) | 7 | 2.10％ | Val(V) | 15 | 4.50％ |
Subsequently, Mn was performed on the screened strains2+The minimum inhibitory concentration test of (4). Using wild type E.coli DH5 alpha and a strain containing empty vector pYES2 (designated as pYES2) as controls and a strain carrying JcVIPP1 gene (designated as JcVIPP1) as an experimental strain, after activating these 3 strains, DH5 alpha, pYES2 and JcVIPP1 were cultured in LB liquid medium and 50mg/l Amp + LB liquid medium, respectively. After overnight culture at 37 ℃ and 200rpm, the bacterial solutions of 3 strains were each diluted to OD6001.00 +/-0.01, and 10 to 10 times5After gradient dilution, in a medium containing 5mM～30mM Mn2+The result of the minimum inhibitory concentration experiment carried out on the LB solid culture medium shows that the JcVIPP1 gene can greatly improve the Mn resistance of the strain2+Ability to stress E.coli at 25mM Mn2+Was grown in the environment of (1).
In FIG. 1, the JcVIPP1 gene improves the Mn resistance of Escherichia coli2+Stress ability. 3 strains are escherichia coli (JcVIPP1) containing vesicle-inducing protein (JcVIPP1) genes in Jatropha curcas plastid 1; a strain containing the empty vector pYES2 (pYES 2); wild type e.coli DH5 α, growth on LB plates at different Mn2+ concentrations, a: 5mM Mn2++ LB solid medium; b: 10mM Mn2++ LB solid medium; c: 15mM Mn2++ LB solid medium; d: 20mM Mn2++ LB solid medium; e: 25mM Mn2++ LB solid medium; f: 30mM Mn2++ LB solid medium.
As shown in FIG. 2, under the condition of normal LB medium culture, wild type Escherichia coli DH5 α, the Escherichia coli recombined with empty vector pYES2 and the Escherichia coli recombined with JcVIPP1 have substantially the same growth conditions, which indicates that the transfer of pYES2 and JcVIPP1 has no obvious influence on the Escherichia coli cultured in LB medium, and the Escherichia coli enters the growth phase from 2h and the platform phase from 18 h.
As can be seen from FIG. 3, at 25mM Mn2+Under the condition of culture medium, wild type Escherichia coli DH5 alpha, Escherichia coli recombined with empty vector pYES2, Escherichia coli recombined with JcVIPP1 are obviously inhibited in growth, the time of entering a growth phase and a plateau phase is obviously delayed, DH5 alpha is most obviously inhibited, weak growth starts from 32h, and after empty vector pYES2 is recombined, 25mM Mn is treated2+The resistance under stress is improved to a certain extent, the growth starts from 28h, and the result shows that the pYES2 vector is transferred to the Escherichia coli at 25mM Mn2+Growth under stress was effected at 25mM Mn after recombination with JcVIPP12+The stress is best in performance, starting from 24h and 4 to the growth phase, and starting from 40h and entering the plateau phase. To exclude vector pYES2 from E.coli at 25mM Mn2+Growth under stress and physiological response, so subsequent experiments chose pYES2 as a control. The performance of each strain is integrated, and the selection determination is carried out at 26hThe enzyme activity was changed at 32h to observe the presence of JcVIPP1 in E.coli at 25mM Mn2+Physiological response in culture.
Influence of CAT Activity under manganese stress of JcVIPP1 Escherichia coli
No obvious difference exists between the CAT enzyme activities of DH5 alpha and pYES2 in the condition that LB culture and 25mM Mn2+ culture are carried out for 26h, and 25mM Mn2+When the culture is carried out for 32h, the CAT enzyme activity of pYES2 is obviously improved compared with that of DH5 alpha (figure 4). Compared with pYES2, the expression of CAT activity of Escherichia coli transferred into JcVIPP1 in LB medium for 26h has very significant effect (p)<0.01) at 25mM Mn2+Both 26h and 32h of culture had significant effects (p)<0.05)。
At 25mM Mn, compared to CAT activity in LB culture2+The CAT activities of DH 5. alpha. and pYES2 were reduced overall in culture (FIG. 4), indicating that CAT expression in the antioxidant system of E.coli was inhibited under manganese stress. The inhibition was counteracted to some extent by the transfer of JcVIPP1, at 25mM Mn2+There was no significant decrease in POD activity in culture. The JcVIPP1 gene is proved to obviously improve the activity of CAT in Escherichia coli. At 25mM Mn, compared to CAT activity in LB culture2+The CAT activity is reduced in culture, which indicates that the expression of CAT in the escherichia coli antioxidant system is inhibited under manganese stress.
Effect of JcVIPP1 on Escherichia coli POD Activity
Whether in LB or 25mM Mn2+The culture medium shows that DH5 alpha has no obvious difference compared with POD enzyme activity of pYES 2. Compared with pYES2, the POD activity expression of JcVIPP1 transferred into 26h of Escherichia coli cultured in LB medium showed significant difference (0.01)<p<0.05) and no significant difference (p) by 32h>0.05) (fig. 5).
At 25mM Mn2+Both 26h and 32h of E.coli in culture had a very significant effect on POD activity expression (p)<0.01) (fig. 5). POD activity in 25mM Mn, as compared with that in LB culture2+Both the POD activities of DH 5. alpha. and pYES2 were reduced in total in culture (FIG. 5), indicating that the expression of POD in the antioxidant system of E.coli was suppressed under manganese stress. The inhibition was counteracted to some extent by the transfer of JcVIPP1, at 25mM Mn2+Under the condition of culture,there was no significant decrease in POD activity.
Effect of JcVIPP1 on Escherichia coli SOD Activity
Whether in LB or 25mM Mn2+There was no significant difference in SOD enzyme activity between cultured ` DH 5. alpha. and pYES2 (FIG. 6). Compared with pYES2, the SOD activity expression of JcVIPP1 transferred to the Escherichia coli cultured in LB medium at 26h and 32h respectively shows very significant difference (p<0.01) and significant difference (p)<0.05) (fig. 6).
At 25mM Mn2+In culture, 26h POD activity expression had no significant effect (p)>0.05), 32h POD activity showed significant difference (p)<0.05) (fig. 6). But at 25mM Mn compared to the SOD activity in LB culture2+The overall SOD activity was significantly increased by about one time under the culture (FIG. 6), which indicates that the expression of SOD in the anti-oxidation system of Escherichia coli was significantly enhanced under the stress of manganese.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (2)
1. A JcVIPP1 recombinant Escherichia coli used as an anti-manganese agent is characterized in that a plastid 1 vesicle inducible protein gene JcVIPP1 in Jatropha curcas is recombined into a vector and is introduced into Escherichia coli to construct the JcVIPP1 recombinant Escherichia coli.
2. A method for constructing the recombinant Escherichia coli JcVIPP1 of claim 1, comprising:
with primer Vipp-F5'-CCG ATGGCCGCAAAATCACAGTTAATT AC-3'; performing RT-PCR on the drought-treated Jatropha curcas cDNA library to obtain a JcVIPP1 full-length coding gene by Vipp-R5'-CCG agcaatcagaattcgTTAGAATTCCTTTCTCTTTTGTCTTAATTCATTG-3', wherein the coding gene has a full length of 1005bp and comprises the following components:
ATGGCCGCAAAATCACAGTTAATTACAGGATTGACCTTGCCATTGCCACCGCCGCATTCTTCCACTTCCTCAACCTCCAATAACAGCAGCAACACTCTCTGTATGGTCAAGCGGCCGCAACTTACGACTTCGTTTTTCAATGGCGGAGTTGAAGCTCTAAAATTTTCTAGGATAAGGACTTGTTCTACTAGGTCCCATTGCTACAGACAAGGTGGAGGTGCTCTTGGCACTCGTATGAATCTTTTTGATCGGTTTGCTAGAGTTGTCAAGTCATATGCAAATGCAATCTTGAGTGGTTTTGAGGACCCGGAAAAAATTCTAGATCAGACGGTTCTTGAAATGAATGATGACTTGACAAAGATGCGTCAGGCCACAGCACAAGTATTGGCATCTCAAAAACGTTTGGAAAATAAATACAAAGCTGCGGAACAAGCTTCTGAGGATTGGTACCGTAAAGCACAACTTGCTCTTCAGAAAGGAGAGGAAGATCTCGCTCGGGAAGCTCTTAAGAGGCGTAAATCTTATGCTGACAATGCAAATTCCTTGAGAGCTCAACTTGATCAACAGAAAAGTGTTGTTGAGAATCTTGTCTCTAATACTCGGCTTTTGGAGAGCAAGATACAGGAGGCAAAGTCTAAAAAAGATACTCTGAAAGCGCGTGCCCAATCTGCAAAAACTCAAACCAAAGTGAATGAGATGCTGGGGAATGTAAATACAAGCAACGCTCTTTCAGCTTTCGAGAAAATGGAAGAGAAAGTATTGCAAATGGAATCAGAAGCTGAAGCACTTGGCCAGTTAGCTACAAGTGAATTGGATGGAAAGTTTGCTTTACTTGAGAGCTCATCTGTTGATGATGATCTTGAGAACCTGAAGAAGGAAATTTCTGGTAGCAAAAAGAGAGGAGAACTGCCGCCCGGTAGAACAGTTGTCAGCAGCTCTGCATTGAGAGATCCTGAGATTGAGATGCAGCTCAATGAATTAAGACAAAAGAGAAAGGAATTCTAA；
encodes a protein consisting of 334 amino acid residues, the amino acid composition is shown in Table 1:
TABLE 1 amino acid composition of JcVIPP1
The product obtained by the amplification of the primers is cut by restriction enzymes Hind III and BamH I, then is connected to a vector pYES2, and is introduced into Escherichia coli DH5 alpha to construct JcVIPP1 recombinant Escherichia coli.
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|---|---|---|---|---|
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Citations (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101203529A (en) * | 2005-02-18 | 2008-06-18 | 诺华疫苗和诊断公司 | Proteins and nucleic acids from meningitis/sepsis-associated escherichia coli |
| US20080193470A1 (en) * | 2005-02-18 | 2008-08-14 | Vega Masignani | Proteins and Nucleic Acids from Meningitis/Sepsis-Associated Escherichia Coli |
| CN101617039A (en) * | 2006-10-20 | 2009-12-30 | 代表亚利桑那大学的亚利桑那校董会 | Modified cyanobacteria |
| US20110124088A1 (en) * | 2009-11-20 | 2011-05-26 | Pei-Fen Yang | Expression vector for expressing recombinant protein in Cyanobacterium |
| AU2012207041A1 (en) * | 2005-02-18 | 2012-08-16 | J. Craig Venter Institute, Inc. | Proteins and nucleic acids from meningitis/sepsis-associated Escherichia Coli |
| CN103571868A (en) * | 2013-11-05 | 2014-02-12 | 深圳大学 | Transgenic regulation and control miRNA (micro ribonucleic acid) gene algae capable of realizing continuous photosynthetic hydrogen production and creation method of gene algae |
| US20150026839A1 (en) * | 2013-07-17 | 2015-01-22 | Monsanto Technology Llc | Maize plants with improved disease resistance |
| CN105001317A (en) * | 2015-08-03 | 2015-10-28 | 中国科学院遗传与发育生物学研究所 | TuVIPP1 protein and encoding gene and application of TuVIPP1 protein |
| CN107475168A (en) * | 2009-09-28 | 2017-12-15 | 全球健康Gsk疫苗学院有限公司 | High foaming Shigella bacterial strain |
| WO2018057879A1 (en) * | 2016-09-23 | 2018-03-29 | Wake Forest University | Modified microorganisms as sustainable sources of omega-3 polyunsaturated fatty acid production |
| CN109750010A (en) * | 2019-03-14 | 2019-05-14 | 中国科学院植物研究所 | A kind of salicornia europaeal laccase and its encoding gene and application |
| US10540572B1 (en) * | 2019-01-31 | 2020-01-21 | StradVision, Inc. | Method for auto-labeling training images for use in deep learning network to analyze images with high precision, and auto-labeling device using the same |
| US20200115705A1 (en) * | 2017-06-06 | 2020-04-16 | Zymergen Inc. | A high-throughput (htp) genomic engineering platform for improving saccharopolyspora spinosa |
| CN111690581A (en) * | 2019-03-15 | 2020-09-22 | 中国科学院微生物研究所 | Method for producing ice nucleoprotein by recombinant escherichia coli fermentation |
| WO2021211990A1 (en) * | 2020-04-17 | 2021-10-21 | Wake Forest University | Lipid compositions and methods of preparation thereof |
-
2021
- 2021-10-13 CN CN202111194678.4A patent/CN113897324B/en active Active
Patent Citations (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101203529A (en) * | 2005-02-18 | 2008-06-18 | 诺华疫苗和诊断公司 | Proteins and nucleic acids from meningitis/sepsis-associated escherichia coli |
| US20080193470A1 (en) * | 2005-02-18 | 2008-08-14 | Vega Masignani | Proteins and Nucleic Acids from Meningitis/Sepsis-Associated Escherichia Coli |
| AU2012207041A1 (en) * | 2005-02-18 | 2012-08-16 | J. Craig Venter Institute, Inc. | Proteins and nucleic acids from meningitis/sepsis-associated Escherichia Coli |
| CN101617039A (en) * | 2006-10-20 | 2009-12-30 | 代表亚利桑那大学的亚利桑那校董会 | Modified cyanobacteria |
| US20110053216A1 (en) * | 2006-10-20 | 2011-03-03 | Vermaas Willem F J | Modified Cyanobacteria |
| CN107475168A (en) * | 2009-09-28 | 2017-12-15 | 全球健康Gsk疫苗学院有限公司 | High foaming Shigella bacterial strain |
| US20110124088A1 (en) * | 2009-11-20 | 2011-05-26 | Pei-Fen Yang | Expression vector for expressing recombinant protein in Cyanobacterium |
| TW201118170A (en) * | 2009-11-20 | 2011-06-01 | Ind Tech Res Inst | Expression vector for expressing recombinant protein in cyanobacterium |
| US20150026839A1 (en) * | 2013-07-17 | 2015-01-22 | Monsanto Technology Llc | Maize plants with improved disease resistance |
| CN103571868A (en) * | 2013-11-05 | 2014-02-12 | 深圳大学 | Transgenic regulation and control miRNA (micro ribonucleic acid) gene algae capable of realizing continuous photosynthetic hydrogen production and creation method of gene algae |
| CN105001317A (en) * | 2015-08-03 | 2015-10-28 | 中国科学院遗传与发育生物学研究所 | TuVIPP1 protein and encoding gene and application of TuVIPP1 protein |
| WO2018057879A1 (en) * | 2016-09-23 | 2018-03-29 | Wake Forest University | Modified microorganisms as sustainable sources of omega-3 polyunsaturated fatty acid production |
| US20200115705A1 (en) * | 2017-06-06 | 2020-04-16 | Zymergen Inc. | A high-throughput (htp) genomic engineering platform for improving saccharopolyspora spinosa |
| US10540572B1 (en) * | 2019-01-31 | 2020-01-21 | StradVision, Inc. | Method for auto-labeling training images for use in deep learning network to analyze images with high precision, and auto-labeling device using the same |
| CN109750010A (en) * | 2019-03-14 | 2019-05-14 | 中国科学院植物研究所 | A kind of salicornia europaeal laccase and its encoding gene and application |
| CN111690581A (en) * | 2019-03-15 | 2020-09-22 | 中国科学院微生物研究所 | Method for producing ice nucleoprotein by recombinant escherichia coli fermentation |
| WO2021211990A1 (en) * | 2020-04-17 | 2021-10-21 | Wake Forest University | Lipid compositions and methods of preparation thereof |
Non-Patent Citations (4)
| Title |
|---|
| LINGANG ZHANG等: "Essential Role of VIPP1 in Chloroplast Envelope Maintenancein Arabidopsis", vol. 24, pages 2 - 3 * |
| NCBI: "PREDICTED: Jatropha curcas membrane-associated 30 kDa protein, chloroplastic(LOC105628294), mRNA", pages 012209692 * |
| 钟罗宝;陈谷;任丹丹;: "集胞藻PCC6803 EGY同源基因破坏突变体的构建及表型分析" * |
| 钟罗宝;陈谷;任丹丹;: "集胞藻PCC6803 EGY同源基因破坏突变体的构建及表型分析", 微生物学报, no. 11, pages 1470 - 1476 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115895939A (en) * | 2022-09-08 | 2023-04-04 | 广西师范大学 | Enterobacter FM-1 and application of remediation microbial inoculum thereof in manganese-polluted environment treatment |
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