CN103966243A - DNA (Deoxyribose Nucleic Acid) molecule and application of DNA molecule in preparing diabetes mouse model - Google Patents
DNA (Deoxyribose Nucleic Acid) molecule and application of DNA molecule in preparing diabetes mouse model Download PDFInfo
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Abstract
The invention discloses a DNA molecule and application of the DNA molecule in preparing a diabetes mouse model. The DNA molecule comprises an expression cassette A and an expression cassette B, wherein the expression cassette A is used to express porcine HSD11B1 gene; the expression cassette B is used to express CHOP gene and hIAPP gene; the porcine HSD11B1 gene is used to encode protein as shown in the sequence 3 in a sequence table; the CHOP gene is used to express protein as shown in the sequence 4 in the sequence table; the hIAPP gene is used to encode protein as shown in the sequence 5 in the sequence table. During use, a recombinant vector can be used to co-transform the three genes in order to induce mouse pancreatic beta cell to start an apoptosis stress related pathway; further the quantity of the mouse pancreatic beta cell is reduced to cause absolute insufficient insulin secretion; with the addition of insulin resistance, caused by HSD11B1, obvious sugar tolerance is generated; finally continuous rise of fasting blood-glucose can be caused to induce diabetes, so that the modeling purpose is achieved.
Description
Technical field
The present invention relates to a kind of DNA molecular and in the application of preparing in Model of diabetic rat.
Background technology
Medical research needs a large amount of animal models relevant to human diseases pathology of setting up.The animal model that can simulate and embody diabetes B pathological change will contribute to people to understand better the essence of disease, provide fundamental basis for prevention and the treatment of diabetes B, the discovery that can be drug target screening, new drug development, novel therapeutic mode provides subjects.
Use genetic engineering technique to prepare the animal model such as mouse and miniature pig, genome to animal model is transformed, can be by proceeding to genes involved, and make it the Method Modeling of up-regulated expression, at insulin resistant and the two approach modeling effort mankind diabetes Bs of defect of insulin secretion, the animal model of preparing than additive method is better.
Summary of the invention
The object of this invention is to provide a kind of DNA molecular and in the application of preparing in Model of diabetic rat.
The invention provides a kind of DNA molecular, comprise expression cassette first and expression cassette second, described expression cassette first is used for expressing pig HSD11B1 gene, and described expression cassette second is used for expressing CHOP gene and hIAPP gene; Protein shown in the sequence 3 of described pig HSD11B1 gene coded sequence table; Protein shown in the sequence 4 of described CHOP expressed sequence table; Protein shown in the sequence 5 of described hIAPP gene coded sequence table.
Described pig HSD11B1 gene specifically can be if the sequence of sequence table 1 be from as shown in 5 ' end 3327-4205 position Nucleotide.Described CHOP gene specifically can be if the sequence of sequence table 1 be from as shown in 5 ' end 7592-8095 position Nucleotide.Described hIAPP gene specifically can be if the sequence of sequence table 1 be from as shown in 5 ' end 8180-8449 position Nucleotide.
In described expression cassette first, can be started by PAPOE promotor the expression of described pig HSD11B1 gene.In described expression cassette second, can be started by PIP promotor the expression of described CHOP gene and described hIAPP gene.
Described PAPOE promotor specifically can be if the sequence of sequence table 1 be from as shown in 5 ' end 1675-3311 position Nucleotide.Described PIP promotor specifically can be if the sequence of sequence table 1 be from as shown in 5 ' end 6102-7591 position Nucleotide.
Described DNA molecular can comprise following element successively to downstream from upstream: insulator, described PAPOE promotor, described pig HSD11B1 gene, BGHpA tailing signal, described insulator, described PIP promotor, described CHOP gene, F-2A sequence, described hIAPP gene, described BGHpA tailing signal; Described insulator is if the sequence 1 of sequence table is from as shown in 5 ' end 7-1674 position Nucleotide; Described PAPOE promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 1675-3311 position Nucleotide; Described BGHpA tailing signal is if the sequence 1 of sequence table is from as shown in 5 ' end 4209-4433 position Nucleotide; Described PIP promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 6102-7591 position Nucleotide; Described F-2A sequence is if the sequence 1 of sequence table is from as shown in 5 ' end 8096-8179 position Nucleotide.
Described DNA molecular specifically can be as follows (a) or (b): (a) sequence 1 of sequence table is from the DNA molecular shown in 5 ' end 7-8674 position Nucleotide; (b) sequence 1 of sequence table is from the DNA molecular shown in 5 ' end 7-8832 position Nucleotide.
Contain described DNA molecular recombinant vectors, expression cassette or transgenic cell line and all belong to protection scope of the present invention.
Described recombinant vectors specifically can be in the multiple clone site of pcDNA3.1 (+) carrier and inserts the recombinant plasmid that described DNA molecular obtains.
Described recombinant vectors specifically can be the small segment (PCMV fragment) between pcDNA3.1 (+) carrier Mul I and Not I restriction enzyme site is substituted by the recombinant plasmid that described DNA molecular obtains.
The present invention also protects above arbitrary described DNA molecular or the application of recombinant vectors, is following (1) or (2) or (3) or (4): (1) makes Model of diabetic rat; (2) bring out mouse generation insulin resistant; (3) for the preparation of the test kit of making Model of diabetic rat; (4) for the preparation of the test kit that brings out mouse generation insulin resistant.
The present invention also protects a kind of reagent, comprises above arbitrary described DNA molecular or recombinant vectors; The purposes of described reagent is following (1) or (2) or (3) or (4): (1) makes Model of diabetic rat; (2) bring out mouse generation insulin resistant; (3) for the preparation of the test kit of making Model of diabetic rat; (4) for the preparation of the test kit that brings out mouse generation insulin resistant.
Described Model of diabetic rat is the mouse that meets following condition: fasting blood sugar higher than >=6.7mmol/L and carbohydrate tolerance test 120min moment blood glucose value >=10.1mmol/L.
The present invention starts HSD11B1 by liver specificity promotor and crosses expression, starting CHOP gene by pancreas specific promoter expresses with the mistake of hIAPP gene and it is consistent with the Space-time speciality of insulin expression (by Furin cracking site with certainly shear polypeptide 2A and be connected the coexpression of realizing two genes, two genes are in second expression cassette, between expression cassette, separate with MAR insulator, insulator intercepts influencing each other of may existing between double-promoter may cause the unsettled drawback of genetic expression simultaneously.When use, can be by a recombinant vectors to three gene cotransformations, also avoided in many plasmids cotransformation system, multichip carrier transformation efficiency problem and Multiple Classes of Antibiotics select and bring loaded down with trivial details), causing that mouse islets β cell starts apoptosis stress related pathways, and then beta Cell of islet quantity is reduced, cause insulin secretion definitely not enough, add the Insulin Resistance that HSD11B1 causes, produce obvious impaired glucose tolerance, finally there is mouse fasting plasma glucose Continued, diabetes occur, reach modeling object.
Brief description of the drawings
Fig. 1 is the structural representation of recombinant plasmid pcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP.
Fig. 2 is in embodiment 2, the relative expression quantity of hIAPP gene in the relative expression quantity of the relative expression quantity of HSD11B1 gene, the interior CHOP gene of pancreas and pancreas in F1 generation children transgenic mice liver in age.
Fig. 3 is in embodiment 2, and F0 is for the relative expression quantity that becomes hIAPP gene in the relative expression quantity of CHOP gene in the relative expression quantity, pancreas of HSD11B1 gene in age transgenic mice liver and pancreas.
Fig. 4 is in embodiment 2, starts timing from the first day of the high sugar of high fat of feeding, the mean body weight of mouse after 0 moment, 1 month, after 2 months and after 3 months.
Fig. 5 is in embodiment 2, starts timing, the result of carbohydrate tolerance test after 0 moment and 2 months from the first day of the high sugar of high fat of feeding.
Fig. 6 is in embodiment 2, mouse fasting blood sugar and carbohydrate tolerance test 120min moment blood glucose value.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment, results averaged for three times.
PcDNA3.1 (+) carrier: Invitrogen, Catalog nos.V790-20.
PEGFP-N1 carrier: BD Biosciences/Clontech, Catalog#6085-1.
Lipofectamine
tM2000 (liposome): Invitrogen.
C57BL/6J mouse: purchased from Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences.
Pair of primers is as follows:
PAPOE-11b F：5'-GCTCCCTTTCCCCCTTAACC-3'；
PAPOE-11b R：5'-AGGCCAAGAAGATCCCCAGA-3'。
Second pair of primer is as follows:
PIP-CHOP F：5'-AGGGAAATGATCCAGAAAGTGC-3'；
PIP-CHOP R：5'-GGACGCAGGGTCAAGAGTAGTG-3'。
The 3rd pair of primer pair is as follows:
CHOP-hIAPP F：5'-GAAACGGAAACAGAGTGG-3'；
CHOP-hIAPP R：5'-GTTGCTGGAATGAACTAAAA-3'。
The structure of embodiment 1, recombinant plasmid pcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP
1, the double chain DNA molecule shown in the sequence 1 of composition sequence table.
In the sequence 1 of sequence table, be the recognition sequence of restriction enzyme Mlu I from 5 ' end 1-6 position Nucleotide, 7-1674 position Nucleotide is MAR insulator (insulator), 1675-3311 position Nucleotide is PAPOE promotor, 3312-4208 position Nucleotide be pig HSD11B1 gene (wherein, 3327-4205 position Nucleotide is open reading frame), 4209-4433 position Nucleotide is BGHpA tailing signal, 4434-6101 position Nucleotide is MARinsulator (insulator), 6102-7591 position Nucleotide is PIP promotor, 7592-8095 position Nucleotide is mouse CHOP gene, 8096-8179 position Nucleotide be F-2A sequence (wherein, 8096-8107 position is the encoding sequence of the amino acid " RAKR " in Furin enzymatic lysis site, 8108-8179 position Nucleotide is that 2A is from the encoding sequence of shearing polypeptide), 8180-8449 position Nucleotide is hIAPP gene (deriving from people's IAPP gene), 8450-8674 position Nucleotide is BGHpA tailing signal, 8833-8840 position Nucleotide is the recognition sequence of restriction enzyme Not I.
2, the double chain DNA molecule obtaining by restriction enzyme Mul I and Not I double digestion step 1, reclaims enzyme and cuts product.
3, with restriction enzyme Mul I and Not I double digestion pcDNA3.1 (+) carrier, reclaim about 4.7kb carrier framework.
4, the carrier framework of the enzyme of step 2 being cut to product and step 3 is connected, and obtains recombinant plasmid
pcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP。Recombinant plasmid
The nucleotide sequence of pcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP is as shown in the sequence 2 of sequence table, and structural representation is shown in Fig. 1.
The Preparation and identification of embodiment 2, transgenic mice
One, the preparation of transgenic mice
1, with restriction enzyme Sca I single endonuclease digestion recombinant plasmid
PcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP (linearizing), obtains linear DNA molecule.
2, the linear DNA molecule that recycling step 1 obtains, be injected in zygote (zygote of the male mouse of the female mouse of C57BL/6J and C57BL/6J), then move into the pregnant mouse of C57BL/6J (acceptor mouse), raise acceptor mouse to its natural production, the offspring rat obtaining is that F0 is for mouse.
3, clip F0 is for the tail of mouse, extract genomic dna, carry out PCR qualification and (adopt respectively following three pairs of primers to carry out PCR qualification: the primer pair of primer pair, CHOP-hIAPP F and the CHOP-hIAPP composition of primer pair, PIP-CHOP F and the PIP-CHOPR composition of PAPOE-11b F and PAPOE-11b R composition; If the result that adopts above-mentioned three pairs of primers to carry out PCR qualification is all positive, mouse to be measured is transgenic mice).
4, get F0 for the public mouse of transgenic mice, every public mouse and 2 female mouse of C57BL/6J carry out natural crossing, female mouse natural production, and the offspring rat obtaining is F1 generation mouse.
5, the tail of clip F1 generation mouse, extracts genomic dna, carries out PCR qualification (primer pair of PCR qualification is with step 3, if the result that adopts three pairs of primers to carry out PCR qualification is all positive, mouse to be measured is transgenic mice).
6, get respectively F1 generation children (7 week age in age, i.e. 1.5 monthly ages) with F0 for becoming (34 week age at age, i.e. 8 monthly ages) liver, the pancreas of transgenic mice (using C57BL/6J mouse as negative control), extracting total RNA reverse transcription is cDNA, taking cDNA as template, taking GAPDH gene as reference gene, identify respectively the relative expression quantity of hIAPP gene in the relative expression quantity of CHOP gene in the relative expression quantity, pancreas of HSD11B1 gene in liver and pancreas by quantitative fluorescent PCR.
Primer pair for the identification of HSD11B1 gene is as follows:
11β35-F：5'-AGACACAGACACGGCCATGA-3'；
11β35-R：5'-TTCGGAGATGGTTGTACGTTGA-3'。
Primer pair for the identification of CHOP gene is as follows:
CHOPzhuzi-F：5'-TGCCTTTCACCTTGGAGACGG-3'；
CHOPzhuzi-R：5'-AGGGCTTTGGGATGTGCGTGT-3'。
Primer pair for the identification of hIAPP gene is as follows:
IAPP75short-F：5'-TGAAAGTCATCAGGTGGAAAAGC-3'；
IAPP75short-R：5'-AGGCGCTGCGTTGCA-3'。
Primer pair for the identification of GAPDH gene is as follows:
GAPDH-F：5'-CATGTTCCAGTATGACTCCACTC-3'；
GAPDH-R：5'-GGCCTCACCCCATTTGATGT-3'。
F1 generation children transgenic mice in age the results are shown in Figure 2.Result shows, in transgenic mice liver, the relative expression quantity of HSD11B1 gene is significantly higher than C57BL/6J mouse, in pancreas of transgenic mouse, the relative expression quantity of CHOP gene is significantly higher than C57BL/6J mouse, and in pancreas of transgenic mouse, the relative expression quantity of hIAPP gene is significantly higher than C57BL/6J mouse.
F0 the results are shown in Figure 3 for one-tenth age transgenic mice.Result shows, in transgenic mice liver, the relative expression quantity of HSD11B1 gene is significantly higher than C57BL/6J mouse, in pancreas of transgenic mouse, the relative expression quantity of CHOP gene is significantly higher than C57BL/6J mouse, and in pancreas of transgenic mouse, the relative expression quantity of hIAPP gene is significantly higher than C57BL/6J mouse.
Above result shows, recombinant plasmid pcDNA3.1-PAPOE-HSD11B1-PIP-CHOP-hIAPP has obtained desirable expression in mouse individual level.
Two, detect and judge insulin resistant by sugar tolerance
Normal feed: DIO diet#D12450K (Research Diets, Inc., New Brunswick, USA).
High-sugar-fat-diet: DIO diet#D12451 (Research Diets, Inc., New Brunswick, USA).
1, packet transaction
First group (Tg HF-HG): adopt normal feed to feed age in male transgenic mice to 13 week of F1 generation, then adopt high-sugar-fat-diet to feed;
Second group (Ng HF-HG): adopt normal feed to feed age in male C 57 BL/6 J mouse to 13 week, then adopt high-sugar-fat-diet to feed;
The 3rd group (Tg Control): adopt normal feed replacement to raise high-sugar-fat-diet and carry out the parallel laboratory test of first group;
The 4th group (Ng Control): adopt normal feed replacement to raise high-sugar-fat-diet and carry out the parallel laboratory test of second group.
2, weight ratio
In step 1, start timing from the first day of the high sugar of high fat of feeding, the mean body weight of mouse is shown in Fig. 4 after 0 moment, 1 month, after 2 months and after 3 months.In Fig. 4, the result of each group is the mean value of 5 public mouse.The result of Fig. 4 shows: the in the situation that of the high sugar induction of high fat, the body weight of transgenic mice is significantly higher than C57BL/6J mouse; In the situation that feeding normal feed, the body weight of transgenic mice is also significantly higher than C57BL/6J mouse.
3, blood sugar tolerance relatively
The method of carbohydrate tolerance test (IPGTT): first make mouse 12h on an empty stomach, then give the D-Glucose (adopting physiological saline as solvent) of 1% (mass ratio) body weight by abdominal injection, taking injection time as starting point, at 0min, 15min, 30min, 45min, 60min, 90min and 120min chronometry mouse tail blood glucose value.
In step 1, start timing from the first day of the high sugar of high fat of feeding, after 0 moment and 2 months, carbohydrate tolerance test the results are shown in Figure 5.
Fig. 5 A is the carbohydrate tolerance test result in 0 moment of packet transaction, and Fig. 5 B is area under the glucose tolerance curve obtaining based on Fig. 5 A (AUC), and under glucose tolerance curve, the larger explanation insulin resistance of area is larger.
Fig. 5 C is the carbohydrate tolerance test result of packet transaction after 2 months, and Fig. 5 D is area under the glucose tolerance curve obtaining based on Fig. 5 C (AUC), and under glucose tolerance curve, the larger explanation insulin resistance of area is larger.
The result of Fig. 5 shows, feeds high-sugar-fat-diet after 2 months, and insulin resistant has significantly occurred transgenosis group mouse, and degree is significantly higher than the high-sugar-fat-diet C57BL/6J same period mouse of feeding.
In Fig. 5, the result of each group is the mean value of 5 public mouse.The result of Fig. 5 shows, the blood glucose value entirety of transgenic mice after high glucose and high fat feed induction hemostasis D-Glucose is higher than control mice, and glucose-tolerant ability declines, and insulin resistant has occurred.
4, fasting blood sugar and carbohydrate tolerance test 120min moment blood glucose value
In step 1, start timing from the first day of the high sugar of high fat of feeding, the fasting blood sugar of transgenic mice after detecting respectively 0 moment, 2 months and after 4 months (is not feed in continuous 12 hours on an empty stomach, detect mouse tail blood sugar), the results are shown in Figure 6A (dotted line represents 6.7mmol/L, and B figure represents 10.1mmol/L).In step 1, start timing from the first day of the high sugar of high fat of feeding, transgenic mice carbohydrate tolerance test 120min moment blood glucose value after recording respectively 0 moment, 2 months and after 4 months (carbohydrate tolerance test 120min moment blood glucose value is called for short 120min moment blood glucose value), method is as follows: first make mouse 12h on an empty stomach, then give the D-Glucose (adopting physiological saline as solvent) of 1% (mass ratio) body weight by abdominal injection, taking injection time as starting point, at 120min chronometry mouse tail blood glucose value, the results are shown in Figure 6B (dotted line represents 10.1mmol/L).
The results are shown in Table 1.
Table 1 fasting blood sugar and carbohydrate tolerance test 120min moment blood glucose value (mmol/L)
Using fasting blood sugar higher than >=6.7mmol/L and carbohydrate tolerance test 120min moment blood glucose value >=10.1mmol/L the judging criterion as diabetes.The high sugar of the high fat of feeding is after 2 months, and Tg HF-HG group can obtain diabetic mice, and Tg Control not yet obtains diabetic mice.
In sum, the transgenic mice that the present invention obtains can be used as diabetic mice, thereby is applied to the research of diabetes mechanism and the exploitation of medicine, has vast application prospect.
Claims (10)
1. a DNA molecular, comprises expression cassette first and expression cassette second, and described expression cassette first is used for expressing pig HSD11B1 gene, and described expression cassette second is used for expressing CHOP gene and hIAPP gene; Protein shown in the sequence 3 of described pig HSD11B1 gene coded sequence table; Protein shown in the sequence 4 of described CHOP expressed sequence table; Protein shown in the sequence 5 of described hIAPP gene coded sequence table.
2. DNA molecular as claimed in claim 1, is characterized in that: described pig HSD11B1 gene is if the sequence 1 of sequence table is from as shown in 5 ' end 3327-4205 position Nucleotide; Described CHOP gene is if the sequence 1 of sequence table is from as shown in 5 ' end 7592-8095 position Nucleotide; Described hIAPP gene is if the sequence 1 of sequence table is from as shown in 5 ' end 8180-8449 position Nucleotide.
3. DNA molecular as claimed in claim 1 or 2, is characterized in that: in described expression cassette first, started the expression of described pig HSD11B1 gene by PAPOE promotor; In described expression cassette second, started the expression of described CHOP gene and described hIAPP gene by PIP promotor.
4. DNA molecular as claimed in claim 3, is characterized in that: described PAPOE promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 1675-3311 position Nucleotide; Described PIP promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 6102-7591 position Nucleotide.
5. DNA molecular as claimed in claim 3, is characterized in that: described DNA molecular comprises following element successively to downstream from upstream: insulator, described PAPOE promotor, described pig HSD11B1 gene, BGHpA tailing signal, described insulator, described PIP promotor, described CHOP gene, F-2A sequence, described hIAPP gene, described BGHpA tailing signal; Described insulator is if the sequence 1 of sequence table is from as shown in 5 ' end 7-1674 position Nucleotide; Described PAPOE promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 1675-3311 position Nucleotide; Described BGHpA tailing signal is if the sequence 1 of sequence table is from as shown in 5 ' end 4209-4433 position Nucleotide; Described PIP promotor is if the sequence 1 of sequence table is from as shown in 5 ' end 6102-7591 position Nucleotide; Described F-2A sequence is if the sequence 1 of sequence table is from as shown in 5 ' end 8096-8179 position Nucleotide.
6. the DNA molecular as shown in claim 5, is characterized in that: described DNA molecular for following (a) or (b): (a) sequence 1 of sequence table is from the DNA molecular shown in 5 ' end 7-8674 position Nucleotide; (b) sequence 1 of sequence table is from the DNA molecular shown in 5 ' end 7-8832 position Nucleotide.
7. contain arbitrary described DNA molecular recombinant vectors, expression cassette or transgenic cell line in claim 1 to 6.
8. recombinant vectors as claimed in claim 7, is characterized in that: described recombinant vectors is for to be substituted by by the small segment between pcDNA3.1 (+) carrier Mul I and Not I restriction enzyme site the recombinant plasmid that described DNA molecular obtains.
9. the application of arbitrary described DNA molecular, recombinant vectors claimed in claim 7 or recombinant vectors claimed in claim 8 in claim 1 to 6, is following (1) or (2) or (3) or (4):
(1) make Model of diabetic rat;
(2) bring out mouse generation insulin resistant;
(3) for the preparation of the test kit of making Model of diabetic rat;
(4) for the preparation of the test kit that brings out mouse generation insulin resistant.
10. a reagent, comprises arbitrary described DNA molecular, recombinant vectors claimed in claim 7 or recombinant vectors claimed in claim 8 in claim 1 to 6; The purposes of described reagent is following (1) or (2) or (3) or (4):
(1) make Model of diabetic rat;
(2) bring out mouse generation insulin resistant;
(3) for the preparation of the test kit of making Model of diabetic rat;
(4) for the preparation of the test kit that brings out mouse generation insulin resistant.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222176A (en) * | 2016-08-08 | 2016-12-14 | 中国农业科学院深圳农业基因组研究所 | A kind of DNA molecular and the application in preparing Model of diabetic rat thereof |
| CN107189986A (en) * | 2017-07-25 | 2017-09-22 | 南京大学 | A kind of reporter gene cell line construction method and its application based on er stress |
| CN110305872A (en) * | 2019-07-17 | 2019-10-08 | 中国农业科学院北京畜牧兽医研究所 | The construction method of miniature pig diabetes B model and application |
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2014
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Non-Patent Citations (3)
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| CHANG-JIANG HUANG等: "High Expression Rates of Human Islet Amyloid Polypeptide Induce Endoplasmic Reticulum Stress–Mediated β-Cell Apoptosis, a Characteristic of Humans With Type 2 but Not Type 1 Diabetes", 《DIABETES》, vol. 56, 31 August 2007 (2007-08-31), pages 2016 - 2027 * |
| DREL VR等: "The leptin-deficient(ob/ob)mouse:a new animal model of peripheral neuropathy of type 2 diabetes and obesity", 《DIABETES》, no. 55, 31 December 2006 (2006-12-31), pages 3335 - 3343 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106222176A (en) * | 2016-08-08 | 2016-12-14 | 中国农业科学院深圳农业基因组研究所 | A kind of DNA molecular and the application in preparing Model of diabetic rat thereof |
| CN107189986A (en) * | 2017-07-25 | 2017-09-22 | 南京大学 | A kind of reporter gene cell line construction method and its application based on er stress |
| CN107189986B (en) * | 2017-07-25 | 2020-08-11 | 南京大学 | Construction method and application of reporter gene cell strain based on endoplasmic reticulum stress |
| CN110305872A (en) * | 2019-07-17 | 2019-10-08 | 中国农业科学院北京畜牧兽医研究所 | The construction method of miniature pig diabetes B model and application |
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