CN105624267A - Method for evaluating Bt protein activation actions of midgut digestive enzymes of insects - Google Patents
Method for evaluating Bt protein activation actions of midgut digestive enzymes of insects Download PDFInfo
- Publication number
- CN105624267A CN105624267A CN201610089908.3A CN201610089908A CN105624267A CN 105624267 A CN105624267 A CN 105624267A CN 201610089908 A CN201610089908 A CN 201610089908A CN 105624267 A CN105624267 A CN 105624267A
- Authority
- CN
- China
- Prior art keywords
- toxin
- activation
- midgut digestive
- digestive enzymes
- insect midgut
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Enzymes And Modification Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides a method for evaluating Bt protein activation actions of midgut digestive enzymes of insects. The method includes proportionally mixing specific digestive enzyme inhibitors and midgut digestive enzyme liquid of the insects with one another; activating Bt protein protoxin in an in-vitro manner; analyzing activation effects by means of polyacrylamide gel electrophoresis; counting results by the aid of software ImageJ 1.46; evaluating the Bt protein activation actions participated by the midgut digestive enzymes by means of variance analysis. The method has the advantages that the method is high in operability and efficiency, is accurate and can be used for evaluating the different digestive enzymes which participate in activation of different types of Bt proteins, primary and secondary actions of the different digestive enzymes during Bt protein activation further can be analyzed, and reference effects can be realized for selecting reasonable Bt composite insecticides or transgenic and gene-stacked crops; the method can be applied to Bt biochemical-level resistance evaluation work.
Description
Technical field
The present invention relates to and a kind of evaluate the insect midgut digestive enzyme method to Bt protein activation effect.
Background technology
Bacillus thuringiensis (Bacillusthuringiensis, Bt) gram-positive bacterium is belonged to, polytype parasporal crystal protein can be produced, i.e. delta-endotoxin or be called insecticidal crystal protein (insecticidalcrystalprotein, ICPs), to Lepidoptera (Lepidoptera), Diptera (Diptera), coleoptera (Coleoptera), Hymenoptera (Hymenoptera), Homoptera (Homoptera), Orthoptera (Orthoptera), the various insects such as Mallophaga (Mallophaga), and nematicide, demodicid mite class and protozoacide etc. have specific insecticidal activity.
The mechanism of action of classical Bt parent toxin is as follows: crystalline protein dissolves in insect midgut, before under the effect of midgut proteinase, toxin is hydrolyzed to activated toxin, after activated toxin is combined with the specific receptor of middle intestinal, subunit oligomerization, formed pore space structure, and it is inserted on middle intestinal teleblem, these holes allow ion and moisture freely to come in and go out cell, cause ion channel or hole, cause cell expansion and cracking, final insect death.
Summary of the invention
It is an object of the invention to provide and a kind of evaluate the insect midgut digestive enzyme method to Bt protein activation effect.
Activation step of the normally off key of right and wrong in Bt action model of Bt toxin, this step is not only related to the Bt toxic effect to insecticide, is likely to the formation participating in resistance simultaneously. conventional research is merely by comparing before and after insect's food-taking or produces the change of enterokinase vigor before and after resistance and carrys out the effect of analyzing proteins enzyme, not yet set up so far and a set of evaluate the various protease method to Bt protein activation effect intuitively, in view of this, the present invention provides a kind of and evaluates the insect midgut digestive enzyme method to Bt protein activation effect, it is specific digestion enzyme inhibitor and insect midgut digestive enzyme liquid to be mixed in proportion, in vitro Bt proteinogen toxin is activated, then pass through polyacrylamide gel electrophoresis activation effect is analyzed, with software I mageJ1.46, result is added up, the Bt protein activation effect that midgut digestive enzymes participates in is evaluated finally by variance analysis.
Insecticide of the present invention includes Lepidoptera, Diptera, coleoptera, Hymenoptera, Homoptera, Orthoptera, Mallophaga insecticide and nematicide, demodicid mite class and protozoacide etc. Preferably, described insecticide is bollworm.
Specific digestion enzyme inhibitor of the present invention is corresponding to specific protease a kind of in insect midgut digestive enzyme, for instance described specific digestion enzyme inhibitor includes the inhibitor N-tosyl-L-lysinechloromethylketone (TLCK) etc. of trypsinlike enzyme.
Specifically, a kind of of the present invention evaluates the insect midgut digestive enzyme method to Bt protein activation effect, comprises the following steps:
1) certain density insect midgut digestive enzyme liquid is prepared;
2) by certain density specific digestion enzyme inhibitor solution and step 1) insect midgut digestive enzyme liquid mix by different proportion, mix with certain density Bt proteinogen toxin soiutions again and hatch a period of time, reaction is terminated with albumen sample-loading buffer, after boiling water boils 5-10min, carry out polyacrylamide gel electrophoresis, coomassie brilliant blue staining;
3) according to coloration result, with software I mageJ1.46, the activation effect of Bt proteinogen toxin is carried out quantitative analysis, finally carry out variance analysis with software SPSS18.0 and evaluate the Bt protein activation effect that midgut digestive enzymes participates in.
Wherein, step 1) the middle insect midgut digestive enzyme liquid prepared, with Bradford standard measure to protein concentration 1mg/mL.
Step 2) in by the specific digestion enzyme inhibitor solution of 1mg/mL and step 1) insect midgut digestive enzyme liquid mix by the volume ratio of 10:1 and 1:1 respectively, gained mixed liquor mixes by the volume ratio of 1:100 with the Bt proteinogen toxin soiutions of 1mg/mL, hatches 5min-2h for 37 DEG C; Then terminate reaction with 5 �� albumen sample-loading buffer, boiling water boils 6min.
Preferably, step 2) in 8% polyacrylamide gel electrophoresis.
Step 2) in set up without there being the insect midgut digestive enzyme liquid of described specific digestion enzyme inhibitor be blank simultaneously, be about to hatch without there being the insecticide of described specific digestion enzyme inhibitor to mix with Bt proteinogen toxin.
The inventive method is workable, precise and high efficiency, can be used for evaluating which different digestive enzyme and take part in the activation of inhomogeneity Bt albumen, different digestive enzyme primary and secondary effect in activated b t albumen can also be analyzed, for selecting rational Bt combine insecticide or turn superposition gene crops and have reference function simultaneously. Additionally, the method can be applicable to the appraisal to Bt biochemistry level resistance.
Accompanying drawing explanation
Fig. 1 is the activation efficiency of trypsinlike enzyme activation Cry1Ac parent toxin and correspondence in the embodiment of the present invention 1; A: swimming lane 1, albumen marker; Swimming lane 2, Cry1Ac parent toxin; Swimming lane 3, trypsinlike enzyme activation Cry1Ac toxin; The trypsinlike enzyme of swimming lane 4,10:1 and TLCK mixed liquor activation Cry1Ac toxin; The trypsinlike enzyme of swimming lane 5,1:1 and TLCK mixed liquor activation Cry1Ac toxin; Swimming lane 3��swimming lane 5, Cry1Ac toxin has been activated 30 minutes; Swimming lane 6, trypsinlike enzyme activation Cry1Ac toxin (blank), the trypsinlike enzyme of swimming lane 7,10:1 and TLCK mixed liquor activation Cry1Ac toxin; The trypsinlike enzyme of swimming lane 8,1:1 and TLCK mixed liquor activation Cry1Ac toxin; Swimming lane 6��swimming lane 8, Cry1Ac toxin has been activated 2 hours. B: the activation efficiency of each swimming lane of swimming lane 3��swimming lane 8 in abscissa 3��8 corresponding A respectively. The digestive efficiency of each time period difference digestion process carries out significance analysis with blank group parent toxin respectively. *: P < 0.05, Independentt-tests, SPSSversion18.0softwareforWindows.
Fig. 2 is the activation efficiency of bollworm midgut digestive enzymes liquid activation Cry1Ac parent toxin and correspondence in the embodiment of the present invention 1; A: swimming lane 1, albumen marker; Swimming lane 2, Cry1Ac parent toxin; Swimming lane 3, midgut digestive enzymes liquid activation Cry1Ac toxin; The midgut digestive enzymes of swimming lane 4,10:1 and TLCK mixed liquor activation Cry1Ac toxin; The midgut digestive enzymes of swimming lane 5,1:1 and TLCK mixed liquor activation Cry1Ac toxin; Swimming lane 3��swimming lane 5, Cry1Ac toxin has been activated 30 minutes; Swimming lane 6, midgut digestive enzymes liquid activation Cry1Ac toxin (blank), the midgut digestive enzymes of swimming lane 7,10:1 and TLCK mixed liquor activation Cry1Ac toxin; The midgut digestive enzymes of swimming lane 8,1:1 and TLCK mixed liquor activation Cry1Ac toxin; Swimming lane 6��swimming lane 8, Cry1Ac toxin has been activated 2 hours. B: the activation efficiency of each swimming lane of swimming lane 3��swimming lane 8 in abscissa 3��8 corresponding A respectively. The digestive efficiency of each time period difference digestion process carries out significance analysis with blank group parent toxin respectively. *: P < 0.05, Independentt-tests, SPSSversion18.0softwareforWindows.
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention. If not specializing, the conventional means that technological means used in embodiment is well known to those skilled in the art, raw materials used it is commercial goods.
Embodiment 1 evaluates the bollworm midgut digestive enzymes method to Bt albumen (Cry1Ac) activation
1,10 5 bollworm in ages (3 secondary pollutants repeat) are prepared, cut epidermis open, take middle intestinal, add appropriate 0.15MNaCl solution, with the abundant homogenate of glass grinding device, then 4 DEG C, 10000g is centrifuged 15min, take supernatant and obtain bollworm midgut digestive enzymes liquid (containing trypsinlike enzyme), utilize Bradford standard measure to protein concentration 1mg/mL.
2, prepare the inhibitor TLCK of trypsinlike enzyme and compare commercial trypsinlike enzyme, with DMSO, inhibitor TLCK is configured to 1mg/mL solution, inhibitor solution and midgut digestive enzymes liquid are pressed respectively the volume ratio mixing of 10:1 and 1:1, as experimental group, and to set up the bollworm midgut digestive enzymes liquid not containing TLCK be blank. Simultaneously with commercial trypsinlike enzyme for matched group, namely compound concentration is the trypsinlike enzyme solution of 1mg/mL, inhibitor solution and trypsinlike enzyme solution are pressed respectively the volume ratio mixing of 10:1 and 1:1, correspondingly, the trypsinlike enzyme solution setting up the 1mg/mL not containing TLCK is blank.
3, with the NaCO of pH10.03Cry1Ac is configured to the solution of 1mg/mL by buffer, and experimental group is to be mixed by the volume ratio of 1:100 with Cry1Ac parent toxin solution by the mixed liquor of ready inhibitor and midgut digestive enzymes liquid, hatches 30 minutes and 2 hours respectively for 37 DEG C; Then terminate reaction with 5 �� albumen sample-loading buffer, boiling water boils 6min. Blank does same treatment.
Matched group is to be mixed by the volume ratio of 1:100 with Cry1Ac parent toxin solution by the mixed liquor of ready inhibitor and trypsinlike enzyme solution, hatches 30 minutes and 2 hours respectively for 37 DEG C; Then terminate reaction with 5 �� albumen sample-loading buffer, boiling water boils 6min. Blank does same treatment.
4, experimental group ready in step 3 and control sample are added to respectively and the polyacrylamide gel of 8% carries out SDS-PAGE electrophoretic separation. After electrophoresis terminates, dye with Coomassie brilliant blue.
5, after dyeing, glue figure is taken pictures, the size depth according to corresponding band, utilize ImageJsoftware (NIH, v1.46) activation effect of Cry1Ac is carried out quantitatively, by the amount (experimental group) of postdigestive Cry1Ac parent toxin divided by the amount of matched group Cry1Ac parent toxin on same glue, it is activation efficiency.
6, evaluate, finally by variance analysis (SPSSversion18.0softwareforWindows), the Cry1Ac activation that midgut digestive enzymes (particularly trypsinlike enzyme) participates in.
In matched group, the activation efficiency of trypsinlike enzyme activation Cry1Ac parent toxin and correspondence is shown in that Fig. 1, Fig. 1 result shows, the trypsinlike enzyme catapepsis (swimming lane 3 and swimming lane 6) that after 30 minutes and 2 hours, Cry1Ac parent toxin can become commercialized. The trypsinlike enzyme of 10:1 and TLCK mixed liquor inhibit the trypsinlike enzyme digestion to Cry1Ac parent toxin all significantly when 30 minutes (P < 0.0001) and 2 hours (P < 0.0001). The efficiency of its activation is 12.06% and 13.04% (swimming lane 4 and swimming lane 7) respectively. The trypsinlike enzyme of 1:1 and TLCK mixed liquor inhibit the trypsinlike enzyme activation to Cry1Ac parent toxin all significantly when 30 minutes (P < 0.0001) and 2 hours (P < 0.0001). The efficiency of its activation is 7.92% and 11.40% (swimming lane 5 and swimming lane 8) respectively. TLCK can effectively suppress the trypsinlike enzyme activation to Cry1Ac parent toxin, and effectively describes the process of this enzyme participation Cry1Ac parent toxin activation.
The activation efficiency of experimental group bollworm midgut digestive enzymes liquid activation Cry1Ac parent toxin and correspondence is shown in Fig. 2. Fig. 2 result shows, when 30 minutes, Cry1Ac parent toxin was digested 87.34% by bollworm midgut digestive enzymes liquid, and within 2 hours, Cry1Ac parent toxin can by midgut digestive enzymes liquid catapepsis (swimming lane 3 and swimming lane 6). When 30 minutes, the midgut digestive enzymes of 10:1 (P=0.037) and 1:1 (P=0.02) and TLCK mixed liquor can suppress the midgut digestive enzymes activation to Cry1Ac parent toxin significantly. The efficiency of its activation is 67.87% and 57.36% (swimming lane 4 and swimming lane 5) respectively. 2 little constantly, the efficiency that Cry1Ac parent toxin activates is 92.66% and 86.67% by the midgut digestive enzymes of 10:1 and 1:1 and TLCK mixed liquor respectively. Result shows that TLCK can effectively suppress the activation to Cry1Ac parent toxin of the bollworm midgut digestive enzymes liquid, describes the trypsinlike enzyme in this midgut digestive enzymes liquid and participates in the process of Cry1Ac parent toxin activation. But after 2 hours, the activation capacity of Cry1Ac parent toxin is not significantly different from (P=0.082 and P=0.097) with midgut digestive enzymes liquid by the midgut digestive enzymes of 10:1 and 1:1 and TLCK mixed liquor, this result shows in midgut digestive enzymes liquid except trypsinlike enzyme participating in activation process, also has other digestive enzyme to take part in Cry1Ac activation simultaneously.
Although, above the present invention is described in detail with a general description of the specific embodiments, but on basis of the present invention, it is possible to it is made some modifications or improvements, and this will be apparent to those skilled in the art. Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (10)
1. evaluate the insect midgut digestive enzyme method to Bt protein activation effect for one kind, it is characterized in that, specific digestion enzyme inhibitor and insect midgut digestive enzyme liquid are mixed in proportion, in vitro Bt proteinogen toxin is activated, then pass through polyacrylamide gel electrophoresis activation effect is analyzed, with software I mageJ1.46, result is added up, evaluate, finally by variance analysis, the Bt protein activation effect that midgut digestive enzymes participates in.
2. method according to claim 1, it is characterised in that described insecticide includes Lepidoptera, Diptera, coleoptera, Hymenoptera, Homoptera, Orthoptera, Mallophaga insecticide and nematicide, demodicid mite class.
3. method according to claim 2, it is characterised in that described insecticide is bollworm.
4. method according to claim 1, it is characterised in that described specific digestion enzyme inhibitor is corresponding to specific protease a kind of in insect midgut digestive enzyme.
5. method according to claim 4, it is characterised in that described specific digestion enzyme inhibitor includes TLCK.
6. method according to claim 5, it is characterised in that comprise the following steps:
1) certain density insect midgut digestive enzyme liquid is prepared;
2) by certain density specific digestion enzyme inhibitor solution and step 1) insect midgut digestive enzyme liquid mix by different proportion, mix with certain density Bt proteinogen toxin soiutions again and hatch a period of time, reaction is terminated with albumen sample-loading buffer, after boiling water boils 5-10min, carry out polyacrylamide gel electrophoresis, coomassie brilliant blue staining;
3) according to coloration result, with software I mageJ1.46, the activation effect of Bt proteinogen toxin is carried out quantitative analysis, finally carry out variance analysis with software SPSS18.0 and evaluate the Bt protein activation effect that midgut digestive enzymes participates in.
7. method according to claim 6, it is characterised in that step 1) the middle insect midgut digestive enzyme liquid prepared, with Bradford standard measure to protein concentration 1mg/mL.
8. method according to claim 6, it is characterized in that, step 2) in by the specific digestion enzyme inhibitor solution of 1mg/mL and step 1) insect midgut digestive enzyme liquid mix by the volume ratio of 10:1 and 1:1 respectively, gained mixed liquor mixes by the volume ratio of 1:100 with the Bt proteinogen toxin soiutions of 1mg/mL, hatches 5min-2h for 37 DEG C; Then terminate reaction with 5 �� albumen sample-loading buffer, boiling water boils 6min.
9. method according to claim 6, it is characterised in that step 2) in 8% polyacrylamide gel electrophoresis.
10. method according to claim 6, it is characterized in that, step 2) in set up without there being the insect midgut digestive enzyme liquid of described specific digestion enzyme inhibitor be blank simultaneously, be about to hatch without there being the insect midgut digestive enzyme liquid of described specific digestion enzyme inhibitor to mix with Bt proteinogen toxin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610089908.3A CN105624267B (en) | 2016-02-17 | 2016-02-17 | Method for evaluating Bt protein activation effect of insect midgut digestive enzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610089908.3A CN105624267B (en) | 2016-02-17 | 2016-02-17 | Method for evaluating Bt protein activation effect of insect midgut digestive enzyme |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105624267A true CN105624267A (en) | 2016-06-01 |
| CN105624267B CN105624267B (en) | 2019-12-24 |
Family
ID=56039609
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610089908.3A Active CN105624267B (en) | 2016-02-17 | 2016-02-17 | Method for evaluating Bt protein activation effect of insect midgut digestive enzyme |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105624267B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103923175A (en) * | 2014-04-17 | 2014-07-16 | 南京农业大学 | Fluorescent substrate for detecting activity of trypsin acting on Cry1A protoxin and application of fluorescent substrate |
-
2016
- 2016-02-17 CN CN201610089908.3A patent/CN105624267B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103923175A (en) * | 2014-04-17 | 2014-07-16 | 南京农业大学 | Fluorescent substrate for detecting activity of trypsin acting on Cry1A protoxin and application of fluorescent substrate |
Non-Patent Citations (4)
| Title |
|---|
| HELICOVERPA ZEA, Y.C. ZHU ET AL: "Interaction of Cry1Ac toxin (Bacillus thuringiensis) and proteinase inhibitors on the growth, development, and midgut proteinase activities of the bollworm", 《PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY》 * |
| JUAN L.JURAT-FUENTES ET AL: "A proteomic approach to study Cry1Ac binding proteins and their alterations in resistant Heliothis virescens larvae", 《JOURNAL OR INVERTEBRATE PATHOLOGY》 * |
| MELANIE FORTIER ET AL: "Effect of Insect Larval Midgut Proteases on the Activity of Bacillus thuringiensis Cry Toxins", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| TETREAU ET AL: "Larval midgut modifications associated with Bti resistance in the yellow fever mosquito using proteomic and transcriptomic approaches", 《BMC GENOMICS》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105624267B (en) | 2019-12-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Melo et al. | Bacillus thuringiensis: mechanism of action, resistance, and new applications: a review | |
| Zhang et al. | Functional analysis of insect molting fluid proteins on the protection and regulation of ecdysis | |
| Rajamohan et al. | Protein engineering of Bacillus thuringiensis δ-endotoxin: Mutations at domain II of CryIAb enhance receptor affinity and toxicity toward gypsy moth larvae | |
| Wang et al. | Angiotensin-converting enzymes modulate aphid–plant interactions | |
| US20110318385A1 (en) | Engineered botulinum neurotoxin c1 with selective substrate specificity | |
| Zimoch et al. | Regulation of chitin synthesis in the larval midgut of Manduca sexta | |
| Pellett et al. | Comparison of the primary rat spinal cord cell (RSC) assay and the mouse bioassay for botulinum neurotoxin type A potency determination | |
| Contreras et al. | Proteome response of Tribolium castaneum larvae to Bacillus thuringiensis toxin producing strains | |
| Coppin et al. | Testing the evolvability of an insect carboxylesterase for the detoxification of synthetic pyrethroid insecticides | |
| Lewandowska et al. | Plant SILAC: stable-isotope labelling with amino acids of Arabidopsis seedlings for quantitative proteomics | |
| Arvidson et al. | Parasitoid Jewel Wasp Mounts Multipronged Neurochemical Attack to Hijack a Host Brain*[S] | |
| Agostoni et al. | Homeostasis of second messenger cyclic-di-AMP is critical for cyanobacterial fitness and acclimation to abiotic stress | |
| Wei et al. | APN1 is a functional receptor of Cry1Ac but not Cry2Ab in Helicoverpa zea | |
| Siddiq et al. | Experimental test and refutation of a classic case of molecular adaptation in Drosophila melanogaster | |
| Pomerantsev et al. | Genome engineering in Bacillus anthracis using tyrosine site-specific recombinases | |
| Gao et al. | Bacterial spore germination receptors are nutrient-gated ion channels | |
| Perera et al. | Generation of a transcriptome in a model lepidopteran pest, Heliothis virescens, using multiple sequencing strategies for profiling midgut gene expression | |
| Sun et al. | Cathepsin O is involved in the innate immune response and metamorphosis of Antheraea pernyi | |
| Molloy et al. | Characterisation of the major extracellular proteases of Stenotrophomonas maltophilia and their effects on pulmonary antiproteases | |
| Lehmann et al. | Novel markers for high-throughput protoplast-based analyses of phytohormone signaling | |
| Quan et al. | Critical domains in the specific binding of radiolabeled Vip3Af insecticidal protein to brush border membrane vesicles from Spodoptera spp. and cultured insect cells | |
| CN105624267A (en) | Method for evaluating Bt protein activation actions of midgut digestive enzymes of insects | |
| Hecht et al. | Inter‐and intramolecular regulation of protein depupylation in Mycobacterium smegmatis | |
| Wang et al. | Archaea is more important than bacteria in driving soil stoichiometry in phosphorus deficient habitats | |
| Castagnola et al. | Alpha-arylphorin is a mitogen in the Heliothis virescens midgut cell secretome upon Cry1Ac intoxication |
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 |