CN107328947B - A kind of evaluation method, restorative procedure and the feed of transgene pig exogenous origin gene integrator position effect and expression product physiological function - Google Patents
A kind of evaluation method, restorative procedure and the feed of transgene pig exogenous origin gene integrator position effect and expression product physiological function Download PDFInfo
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- CN107328947B CN107328947B CN201710728980.0A CN201710728980A CN107328947B CN 107328947 B CN107328947 B CN 107328947B CN 201710728980 A CN201710728980 A CN 201710728980A CN 107328947 B CN107328947 B CN 107328947B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/174—Vitamins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
Abstract
The invention discloses a kind of evaluation methods of transgene pig exogenous origin gene integrator position effect and expression product physiological function, are related to transgene pig detection field.The evaluation method of transgene pig exogenous origin gene integrator position effect and expression product physiological function disclosed by the invention comprising：Detection turns the blood physiological and biochemical indices and urine physiological and biochemical index of 1 gene pigs of fat.The evaluation method can be to transgene pig exogenous origin gene integrator position effect and expression product to the growth performance of transgene pig influence accurately and reliably evaluated, help to accurately control 1 gene expressions of fat in turning 1 gene pig breeding process of fat.
Description
Technical field
The present invention relates to transgene pig detection fields, in particular to a kind of transgene pig exogenous origin gene integrator position
The evaluation method of effect and expression product physiological function, restorative procedure and feed.
Background technology
The ratio that total fatty and saturated fat intake accounts for total amount of heat in diet is constantly reduced, ω -6 aliphatic acid (ω -
6PUFAs, polyunsaturated fatty acids) intake increases and omega-fatty acid (ω -3PUFAs) is reduced, it can cause
ω -6 and ω -3 ratios reach 18:1 or more, such case, which has angiocardiopathy, chronic disease, to be significantly affected, and can be made
Overweight and fat illness rate dramatically increases.
ω -6 aliphatic acid and ω -6 improve fat risk, red blood cell with ω -3 ratios in red blood cell (RBC) membrane phospholipid
Membrane phospholipid high level ω -3 reduces fat risk.Eicosapentaenoic acid (EPA) and docosahexaenoic acid are taken in by increase
(DHA) disease generation can be reduced, ω -6/ ω -3 ratio balances are of great significance to health and fat prevention.
ω -3PUFAs can reduce cholesterol level in film fat, increase the mobility of cell membrane, reduce what cardiovascular and cerebrovascular disease generated
Ratio has the effects that anti-inflammatory, anticoagulation, inhibits tumour growth, influences a variety of human nerve's property diseases.
People and mammal lack polyunsaturated fatty acid dehydrogenase, can not generate ω -6 and ω -3PUFAs, two kinds of objects
Matter must be obtained from food.Caenorhabditis elegan Fat-1 (Fatty acid desaturases, ω -3 polyunsaturated fatty acids
Dehydrogenase) ω -3 polyunsaturated fatty acids dehydrogenases of gene code have ω -6PUFAs being converted into ω -3PUFAs'
Ability.Using transgenic animals method production long-chain omega -3PUFAs be solve the effective ways that are short of food of ω -3PUFAs it
One, turn fat-1 gene pigs and improve pork nutritive value, there is good application prospect.
Turn fat-1 gene pig breedings to need to use PCR and Southern hybridization blot assays primary to turning fat-1 genes
And passage production piglet is detected, and determines integration of the fat-1 genes in pig genome.Turn fat-1 gene pig external sources
Fat-1 genes whether effective expression, expression product can reach expected physiologic effect and gene integration expression to transgene pig
It is the important test content of transgene pig breeding and realization business application with health either with or without harm.
Invention content
The purpose of the present invention is to provide a kind of transgene pig exogenous origin gene integrator position effects and expression product physiology work(
The evaluation method of energy, which can be to transgene pig exogenous origin gene integrator position effect and expression product to transgene pig
The influence of growth performance accurately and reliably evaluated, contribute in turning fat-1 gene pig breeding process to accurately control
Fat-1 gene expressions.
Another object of the present invention is to provide a kind of restorative procedure of transgene pig gene integration position effect side effect,
The restorative procedure can eliminate the side effect that transgene pig gene integration position effect brings transgene pig growth and development, restore to turn
The normal physiological function of gene pig damaged organ.
Another object of the present invention is to provide a kind of for repairing transgene pig gene integration position effect side effect
Feed after transgene pig eats the feed, can eliminate the side effect that gene integration position effect brings growth and development.
The invention is realized in this way：
A kind of evaluation method of transgene pig exogenous origin gene integrator position effect and expression product physiological function comprising：
Detection turns the blood physiological and biochemical indices and urine physiological and biochemical index of fat-1 gene pigs.
A kind of restorative procedure of transgene pig gene integration position effect side effect comprising：
F1 generation transgene pig is hybridized with feminine gender production pig, obtains F2 for transgenic positive pig；
The feed of the compound additive containing safflower seed oil is fed for transgenic positive pig to the F2 of acquisition.
It is a kind of for repairing the feed of transgene pig gene integration position effect side effect, add containing safflower seed oil is compound
The agent, the safflower seed oil compound additive is added to include：Edible safflower-seed oil, fish meal and vitamine supplement.
The invention has the advantages that：
The evaluation method of transgene pig exogenous origin gene integrator position effect and expression product physiological function provided by the invention,
It is counted by blood physiological and biochemical indices and the urine physiological and biochemical index etc. that detect transgene pig (such as turning fat-1 gene pigs)
According to establishing physiological and biochemical index and reproductive performance, pig growth and development situation association analysis method, judgement turns fat-1 gene pig bases
Because integrating position effect, the operation of specification test experience contributes to more completely, reliably and accurately to the growth of transgene pig
It can be judged with developmental state；
In addition, the restorative procedure of transgene pig gene integration position effect side effect provided by the invention, by by F1 generation
Transgene pig (such as F1 generation turns fat-1 gene pigs) hybridizes with feminine gender production pig, obtains F2 for transgenic positive pig, and feed and contain
The feed of safflower seed oil compound additive eliminates exogenous origin gene integrator position effect side effect.The restorative procedure, which can be eliminated, turns base
Because of the side effect that pig (such as turning fat-1 gene pigs) gene integration position effect brings transgene pig growth and development, restore to turn
The normal physiological function of gene pig damaged organ provides reliable material to turn fat-1 gene pig breeding modified test methods, increases
Transgene pig meat edible safety, beneficial to the health for ensureing people.
In addition, the feed provided by the present invention for repairing transgene pig gene integration position effect side effect, contains
Safflower seed oil compound additive, safflower seed oil compound additive include：Edible safflower-seed oil, fish meal and vitamine supplement.
After transgene pig (such as turning fat-1 gene pigs) is edible, the secondary work that gene integration position effect brings growth and development can be eliminated
With physiology work(of the recovery transgenosis pig (such as turning fat-1 gene pigs) due to damaged organ caused by gene integration position effect
Can, render transgenic pig normal growth improves its disease resistance.
Description of the drawings
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the PCR qualification results to turning fat-1 gene pigs that the embodiment of the present invention 1 provides；
Fig. 2 is the Southern blot analysis results to turning fat-1 gene pigs that the embodiment of the present invention 1 provides.
Specific implementation mode
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Below to a kind of transgene pig exogenous origin gene integrator position effect of the embodiment of the present invention and expression product physiology work(
Evaluation method, restorative procedure and the feed of energy are specifically described.
ω -3 polyunsaturated fatty acids (PUFAs) are in a variety of diseases such as prevention and treatment angiocardiopathy, arthritis, cancers
Sick aspect plays an important role, and participates in a variety of pathological processes, reduces the generation of angiocardiopathy, has to tumour and inhibits to make
With.Fat-1 genes are imported in pig genome using transgenic technology, are improved ω -3 polyunsaturated fatty acids in pork and are contained
Amount, is beneficial to the health of people, the cultivation for turning fat-1 gene pigs has important research meaning in terms of improving pork nutritive value
And application prospect.
There are many ω -3 PUFAs physiological functions, and amount acts on unobvious less, and amount crosses conference and inhibits immunity, damage
Cell membrane function promotes tumour, vascular diseases development.Needing increase detection project evaluation to turn fat-1 gene pig foreign genes is
No suitable high efficient expression and expected physiological function is obtained, the physiological function for the ω -3PUFAs that various aspects evaluation endogenous generates.
It is presently used for judging that the material for turning fat-1 gene pig gene integration position effects only has liver dysfunction detection report
Road, it is therefore desirable to increase detection project and clinical data, accurate judgement turns fat-1 gene pig gene integration position effects to pig
The influence degree of renal function, angiocardiopathy, pipe intestinal digesting function.Reasonable association analysis method is established, while needing specification
Testing cost is saved in operation, improves working efficiency.
Based on this, on the one hand, the present invention provides transgene pig exogenous origin gene integrator position effects and expression product physiology
The evaluation method of function comprising：Detection turns the blood physiological and biochemical indices and urine physiological and biochemical index of fat-1 gene pigs.
Further, in some embodiments of the present invention, the blood physiological and biochemical indices include：Lactic acid (LD),
Glutamic-pyruvic transaminase (ALT), lactic dehydrogenase (LDH), cholinesterase (CHE), total protein (TP), albumin (ALB), globulin
(GLO), blood glucose (GLU), high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C).
Further, in some embodiments of the present invention, the urine physiological and biochemical index includes：Bilirubin
(BIL), urobilinogen (UBG), ketoboidies (KET), vitamin C (VITC), glucose in urine (Glu), Urine proteins (Pro), urine occult blood (BLD),
PH, nitrite (NIT), leucocyte (Leu) and proportion (SG).
Further, in some embodiments of the present invention, the transgene pig is to turn fat-1 gene pigs.
It should be noted that the evaluation method of the present invention is not limited only to turn fat-1 gene pigs, it can also be used to evaluate other kinds
The health status and gene integration position effect of class transgene pig.
The evaluation method of transgene pig exogenous origin gene integrator position effect and expression product physiological function provided by the invention,
It is firstly introduced urine physiological and biochemical index, the blood physiology and biochemistry by detecting transgene pig (such as turning fat-1 gene pigs) refers to
Mark (such as lactic acid, glutamic-pyruvic transaminase, lactic dehydrogenase, cholinesterase, total protein, albumin, globulin, blood glucose, high density fat
Protein cholesterol and low density lipoprotein cholesterol) and urine physiological and biochemical index (bilirubin, urobilinogen, ketoboidies, vitamin C,
Glucose in urine, Urine proteins, urine occult blood, pH, nitrite, leucocyte and proportion.) etc. data, establish physiological and biochemical index with breeding
Performance, pig growth and development situation association analysis method, judgement turn fat-1 gene pig gene integration position effects, and specification detection is real
Operation is tested, contribute to more complete, reliable and accurately judges to turn fat-1 gene pig gene integration position effects to pig kidney function
The influence degree of energy, angiocardiopathy, pipe intestinal digesting function and liver function.
On the other hand, the present invention provides a kind of restorative procedure of transgene pig gene integration position effect side effect,
Including：
F1 generation transgene pig is hybridized with feminine gender production pig, obtains F2 for transgenic positive pig；
The feed of the compound additive containing safflower seed oil is fed for transgenic positive pig to the F2 of acquisition.
Further, in some embodiments of the present invention, the safflower seed oil compound additive includes：Kardiseed is eaten
With oil, fish meal and vitamine supplement.
Further, in some embodiments of the present invention, the feed contains based on 100kg：Edible safflower-seed oil
3-3.5kg, fish meal 3-3.5kg and vitamine supplement 7-9g.
Further, in some embodiments of the present invention, the transgene pig is to turn fat-1 gene pigs.
The restorative procedure of transgene pig gene integration position effect side effect provided by the invention, by by F1 generation transgenosis
Pig (such as F1 generation turns fat-1 gene pigs) hybridizes with feminine gender production pig, obtains F2 for transgenic positive pig, and feed and contain kardiseed
The feed of oily compound additive eliminates exogenous origin gene integrator position effect side effects of the F2 for transgene pig.The restorative procedure can
Eliminate the secondary work that transgene pig (such as turning fat-1 gene pigs) gene integration position effect brings transgene pig growth and development
It, can to turn that fat-1 gene pig breeding modified test methods provide with, the normal physiological function of recovery transgenosis pig damaged organ
By material, increase transgene pig meat edible safety, beneficial to the health for ensureing people.
In another aspect, the present invention provides a kind of feedings for repairing transgene pig gene integration position effect side effect
Material, containing safflower seed oil compound additive, the safflower seed oil compound additive includes：Edible safflower-seed oil, fish meal and
Vitamine supplement.
Further, in some embodiments of the present invention, the feed contains based on 100kg：Edible safflower-seed oil
3-3.5kg, fish meal 3-3.5kg and vitamine supplement 7-9g.
Further, in some embodiments of the present invention, the feed contains based on 100kg：Edible safflower-seed oil
3.2kg, fish meal 3.34kg and vitamine supplement 0.008kg.
Further, in some embodiments of the present invention, the transgene pig is to turn fat-1 gene pigs.
Provided by the present invention for repairing the feed of transgene pig gene integration position effect side effect, contain kardiseed
Oily compound additive, safflower seed oil compound additive include：Edible safflower-seed oil, fish meal and vitamine supplement.Transgenosis
After pig (such as turning fat-1 gene pigs) is edible, the side effect that gene integration position effect brings growth and development can be eliminated, it is extensive
Multiple transgene pig (such as turning fat-1 gene pigs) is made due to the physiological function of damaged organ caused by gene integration position effect
Transgene pig normal growth improves its disease resistance.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
The evaluation side of transgene pig exogenous origin gene integrator position effect and expression product physiological function provided in this embodiment
Method is as follows：
1 turn of fat-1 gene pig exogenous origin gene integrators detection
1.1 turn fat-1 gene pigs using the detection of PCR methods
Ear tissue sample is acquired to production piglet (F1 generation turns fat-1 gene pigs), with Life Technologies, Inc. of U.S. genome
Genome DNA extraction kit extracts total DNA.Each sample takes DNA 120ng, and PCR amplification is carried out using primer HFP1 and HFP2
HFat-1 genetic fragments.Specific amplified fragment length is 600bp, and detection hFat-1 genes are integrated in pig genome.
Amplification uses primer：
HFP1:5’-GTGATGCTATTGCTTTATTTGTAAC-3’；
HFP2:5’-GGAGCAGTGGTGGAATGCCT-3’
PCR reaction systems：2xPCR reaction mixture 12.5uL, DNA sample 120ng, 1uL 10umol/L primer HFP1,
1uL 10umol/L primer HFP2, add ddH2O to total volume 25uL.Test reaction condition：94℃2min；94℃30s,62℃
40s, 72 DEG C of 45s, 30 circular responses；72 DEG C of extension 5min.2% agarose gel electrophoresis checks amplification.
The results are shown in Figure 1 (in figure：1 represents positive control, and 2 represent negative control, 3-16 representative samples, M representation DNAs
Molecular labeling), 3,7,8,9,10,13,15, No. 16 samples have band in the position of 600bp, turn fat- for what PCR methods detected
1 gene masculine pig.
1.2 Southern Blot Hybridization Analysis fat-1 gene integrations
What PCR methods detected turns fat-1 gene pigs, and phenol extraction method purifies DNA sample, uses Southern traces
Hybridization check fat-1 gene integration situations.With 37 DEG C of digestion 7h of 10uL (100IU) restriction enzymes XbaI, sample digestion
It vacuumizes and is concentrated to 15uL.Detection is completed using Roche Holding Ag's digoxigenin labeled kit for detecting nucleic acid.PCR amplification fat-1 bases
Because of 600bp segments, recovery purifying 600bp amplified fragments product is as Southern blot hybridization probes.Positive pig specific hybridization
Band is 1.8Kb, and negative pig does not have hybrid belt.
The results are shown in Figure 2 (in figure：1 is positive control, and 2 negative controls, 3-6 is sample), 3,4, No. 6 samples are
The detection of Southern blot hybridizations turns fat-1 gene masculine pigs.
In 2F1 generations, turn the detection of fat-1 gene pigs physiological and biochemical index, reproductive performance, pig growth and development situation measurement
Blood physiological and biochemical indices (the breast that 20 F1 generations of detection turn fat-1 gene pigs is detected with automatic clinical chemistry analyzer
Sour (LD), glutamic-pyruvic transaminase (ALT), lactic dehydrogenase (LDH), cholinesterase (CHE), total protein (TP), albumin (ALB),
Globulin (GLO), blood glucose (GLU), high-density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C)),
Its urine physiological and biochemical index (bilirubin (BIL), urobilinogen (UBG), ketoboidies is detected with urine automabic analyzer (German section is precious)
(KET), vitamin C (VITC), glucose in urine (Glu), Urine proteins (Pro), urine occult blood (BLD), pH, nitrite (NIT), leucocyte
(Leu) and proportion (SG)), judgement turn fat-1 gene pig gene integration position effects to pig renal function, angiocardiopathy, stomach
The influence degree of enteron aisle digestive function and liver function, to produce field feminine gender pig as a contrast.
As a result it shows：F1 generation turns the detection of fat-1 gene pigs physiological and biochemical index, reproductive performance, the survey of pig growth and development situation
Show the liver function of partial transgenic positive pig by minor injury surely.Compared with production field negative test control pig, it is damaged
Pig proportional band is more than 25~50%, illustrates that this damage is caused by foreign gene imports.F1 generation turns this of fat-1 gene pigs
Disease is derived from the genetic disease of parental generation positive boar, statistics indicate that slightly decline is also parental generation positive boar reproductive performance
Caused by same cause being imported by foreign gene.Integration of the fat-1 genes in pig genome affects the expression of other genes,
It produces and integrates position effect side effect.
In 3F2 generations, turn the detection of fat-1 gene pigs physiological and biochemical index, reproductive performance, pig growth and development situation measurement
3.1 blood physiological and biochemical indices detect
Turn fat-1 gene pigs blood plasma lactic acid, glutamic-pyruvic transaminase, total protein, blood with automatic clinical chemistry analyzer detection F2 generations
Sugar, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, total cholesterol, albumin, lactic dehydrogenase, cholinesterase
Blood physiological and biochemical indices, wherein glutamic-pyruvic transaminase 1 are less than normal value, and 2, blood glucose is less than normal value, and total cholesterol 5 is low
In normal value；2 head height of high-density lipoprotein cholesterol is in normal value, 3 head height of low density lipoprotein cholesterol in normal value, lactic acid
Dehydrogenase 2 head height is normal in normal value, other indexs.A small number of indexs are abnormal in blood parameters is drawn by disease infection
It rises, there is no difference between transgenic positive pig and negative pig.5 pigs of total cholesterol show there are 5 less than normal value
The physiologic effect that transgenic positive pig gene expression has obtained.In F2 generations, turn fat-1 gene pigs renal function, angiocardiopathy, stomach and intestine
Road digestive function, liver function are normal.
3.2 urine physiological and biochemical indexs detect
23 first-born production field feminine gender pigs, 5 F2 are acquired for fat-1 gene masculine pig freshly voided urines are turned, use urine automatic analyzer
It is instrument (German section precious) test bilirubin, urobilinogen, ketoboidies, vitamin C, glucose in urine, Urine proteins, urine occult blood, PH, nitrite, white
Cell, proportion totally 11 urine physiological and biochemical indexs, as a result see the table below shown in 1.
Table 1
The testing result of table 1 shows：Negative pig has 6 projects to show as the positive, is respectively：Vitamin C is 43%, urinates egg
It is 35%, pH 50%, nitrite 10%, leucocyte 5%, proportion 17% in vain；And in F2 generations, turn fat-1 gene masculines
Pig only has 2 projects and shows as the positive, is respectively：Urine proteins are 20%, pH 60%.In F2 generations, turn fat-1 gene masculine pigs urine
Liquid physiological and biochemical index masculine item quantity and ratio illustrate that F2 generations turn fat-1 gene masculines significantly lower than production field feminine gender pig
The urinary system health status of pig has clear improvement.
Comparative example 1
To turning the test of fat-1 gene pig blood physiological and biochemical indices and Gernral Check-up
21 negative tests compare pigs and 20 turn 15 blood physiological and biochemical indices test datas of fat-1 gene masculines pig
It see the table below, completed using automatic clinical chemistry analyzer, as a result as shown in table 2.
Table 2
The statistical data from table 2 is it is found that have 4 positive pig lactic acid concns to increase, under 2 positive pig gpt levels
Drop.10 positive pig high-density lipoprotein cholesterol contents are less than normal pig, 1 positive pig low density lipoprotein cholesterol content
Less than normal pig.5 positive pig lactic dehydrogenases are dense to be higher than normal value, and high-density lipoprotein cholesterol concentration is less than normal
Value.5 positive pig gamma globulins are horizontal normal relatively low, and 4 positive pig cholinesterase are normally relatively low, and other indexs are all normal.
Physiological and biochemical index test result shows the liver function of partial transgenic positive pig by minor injury.With production field
Negative test control pig is compared, and it is more than 25~50% to be damaged pig proportional band, illustrates that this damage is that foreign gene importing causes
's.This disease that F1 generation turns fat-1 gene pigs is derived from the genetic disease of parental generation positive boar, statistics indicate that parental generation is positive
Property boar reproductive performance slightly to decline be also caused by importing same cause by foreign gene.
To sum up, it is logical to turn influence of the fat-1 gene pig gene integration position effects to transgene pig growth and development for existing evaluation
Blood physiological and biochemical indices are often only detected, do not detect urine physiological and biochemical index, therefore this evaluation method has limitation, from
Influence of the gene integration position effect to liver function can only be judged in data, cannot completely, comprehensively judge gene integration position
Set influence of the effect to other organs.
And the transgene pig exogenous origin gene integrator position effect and expression product physiological function that the embodiment of the present invention 1 provides
Evaluation method not only detects blood physiological and biochemical indices, also introduces urine physiological and biochemical index, therefore, is given birth to by urine physiology
Change index as a result, can also judge influence degree of the gene integration position effect to pig urinary system, and then more comprehensively
Completely influence of the evaluation transgene pig exogenous origin gene integrator position effect to transgene pig growth and development.
Therefore, pass through the blood physiological and biochemical indices and urine physiology of detection transgene pig (such as turning fat-1 gene pigs)
The data such as biochemical indicator establish physiological and biochemical index and reproductive performance, pig growth and development situation association analysis method, judge to turn
Fat-1 gene pig gene integration position effects, the operation of specification test experience contribute to more completely, reliably and accurately to turning
The growth performance and developmental state of gene pig judge.
Embodiment 2
Present embodiments provide the restorative procedure for turning fat-1 gene pig gene integration position effect side effects comprising：With
Feed feeding containing safflower seed oil compound additive turns fat-1 gene pigs, and safflower seed oil compound additive is eaten by kardiseed
Oil, fish meal and vitamine supplement composition.
Wherein, feeds utilized based on 100kg, contain：Full price fattening pannage 93.452kg (is just reached purchased from Wuhan and is raised
Expect Co., Ltd), edible safflower-seed oil 3.2kg, fish meal 3.34kg and vitamine supplement 0.008kg (be purchased from Wuhan multidimensional
Scientific and technological feed corporation,Ltd).
Full price fattening pannage just reaches feed corporation,Ltd purchased from Wuhan, and multidimensional compound additive is purchased from Wuhan multidimensional science and technology
Feed corporation,Ltd, various vitamin contents are shown in product description.
Embodiment 3
By the restorative procedure that embodiment 2 provides, fat-1 gene masculine pigs are turned to the F2 generations in embodiment 1 and are fed, are raised
After feeding, gene integration position effect side effect is judged by detecting blood physiological and biochemical indices and urine physiological and biochemical index
Repairing effect.
In 1F2 generations, turn the front and back blood physiological and biochemical indices detection of fat-1 gene pigs feeding
15 positive Swine plasma lactic acid (LD), glutamic-pyruvic transaminase (ALT), total proteins are detected with automatic clinical chemistry analyzer
(TP), blood glucose (GLU), high-density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total cholesterol
(TC), albumin (ALB), lactic dehydrogenase (LDH), cholinesterase (CHE), triglyceride (TG), tumor necrosis factor-alpha
(TNF-α), interleukin 12 (IL-12) totally ten three indexs, the results are shown in Table 3.
Table 3
1 pig glutamic-pyruvic transaminase is less than normal value after feeding experiment, and 2 pigs of blood glucose are less than normal value, 5 pig total cholesterols
Less than normal value (close to normal minimum).2 head height of high-density lipoprotein cholesterol is in normal value, low density lipoprotein cholesterol
3 head heights are higher than normal value in normal value, 2 pigs of lactic dehydrogenase, and other indexs are normal.A small number of indexs in blood parameters
Abnormal is not have difference between transgenic positive pig and negative pig caused by being infected by disease.5 pigs of total cholesterol
Show the physiologic effect for there are 5 transgenic positive pig gene expressions to obtain (close to normal minimum) less than normal value.15
Pig total cholesterol average value is less than test pig before feeding, and horizontal in normal low value, F2 generations turn fat-1 gene pigs renal function, the heart
Vascular diseases, pipe intestinal digesting function, liver function are normal.5 pig TNF-α concentration normal value low values, 6 pig IL-12 are dense
Normal value high level is spent, partial transgenic pig disease resistance is improved.Safflower seed oil compound additive can improve transgene pig
Constitution restores test pig organ normal physiological function.
In 2F2 generations, turn the front and back urine physiological and biochemical index detection of fat-1 gene pigs feeding
Acquire 23 first-born production field feminine gender pigs, 5 external form constitutions it is poor turn fat-1 genes F2 generations positive pig freshly voided urine,
With German section's treasured urine automabic analyzer test bilirubin, urobilinogen, ketoboidies, vitamin C, glucose in urine, Urine proteins, urine occult blood, PH,
Nitrite, leucocyte, proportion totally 11 physiological and biochemical indexs.As a result as shown in table 4 (before feeding), table 5 (after feeding).
Table 4
Table 5
Testing result shows：Using the compound additive containing safflower seed oil feed feeding before, as shown in table 4, negative pig with
It is close to turn fat-1 gene pig urcine physiological and biochemical index positive indication data, two groups of test pig renal functions, digestion and absorption function feelings
Condition is essentially identical.
After the feed feeding of the compound additive containing safflower seed oil, as shown in table 5, negative pig has 6 projects to show as
The positive is respectively：Vitamin C is 43%, Urine proteins 35%, pH 50%, nitrite 10%, leucocyte 5%,
Proportion is 17%；F2 generations, which turn fat-1 gene masculine pigs, has 2 projects to show as the positive, is respectively：Urine proteins are 20%, pH
It is 60%.In F2 generations, turn fat-1 gene masculine pig urcine physiological and biochemical index masculine item quantity and ratio significantly lower than production field
Negative pig illustrates that F2 has clear improvement for the urinary system health status for turning fat-1 gene masculine pigs.
The front and back negative pig urcine physiological and biochemical index positive indication data of feeding are close, and safflower seed oil compound additive is to the moon
Property pig urinary system health, abilities of digestive and absorption do not improve function.And the feed of the compound additive containing safflower seed oil is used to raise
F2 is fed for fat-1 gene pigs are turned, pig blood urine physiological and biochemical index testing result shows using the compound addition of the safflower seed oil
The physiological health side effect that gene integration position effect can be overcome to generate after agent restores pig normal physiological function, and it is anti-to improve pig
Sick ability.
Embodiment 4
The present embodiment provides a kind of feeds for repairing transgene pig gene integration position effect side effect, containing red
Flower seed oil compound additive, the safflower seed oil compound additive include：Edible safflower-seed oil, fish meal and vitamine supplement.
With ground, which contains based on 100kg：Edible safflower-seed oil 3.2kg, fish meal 3.34kg and vitamin add
Add agent 0.008kg, remaining group is divided into all-sided fertilizer and educates pannage.Edible safflower-seed oil 3.2kg, fish meal 3.34kg and vitamin add
Agent 0.008kg is added to form the safflower seed oil compound additive.
It can be repaired using feed provided in this embodiment and turn fat-1 gene pig gene integration position effect side effects, it is extensive
Multiple pig normal physiological function, improves pig disease resistance (effect is as described in Example 3).
Embodiment 5
The present embodiment provides a kind of feeds for repairing transgene pig gene integration position effect side effect, containing red
Flower seed oil compound additive, the safflower seed oil compound additive include：Edible safflower-seed oil, fish meal and vitamin addition
Agent.
The feed contains based on 100kg：Edible safflower-seed oil 3kg, fish meal 3kg and vitamine supplement 7g.
Effect provided in this embodiment for repairing the feed of transgene pig gene integration position effect side effect is the same as real
Apply example 4.
Embodiment 6
The present embodiment provides a kind of feeds for repairing transgene pig gene integration position effect side effect, containing red
Flower seed oil compound additive, the safflower seed oil compound additive include：Edible safflower-seed oil, fish meal and vitamine supplement.
The feed contains based on 100kg：Edible safflower-seed oil 3.5kg, fish meal 3.5kg and vitamine supplement 9g；Its
Remaining group is divided into all-sided fertilizer and educates pannage.
Effect provided in this embodiment for repairing the feed of transgene pig gene integration position effect side effect is the same as real
Apply example 4.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, any made by repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (3)
1. a kind of restorative procedure of transgene pig gene integration position effect side effect, which is characterized in that it includes：
F1 generation transgene pig is hybridized with feminine gender production pig, obtains F2 for transgenic positive pig；
The compound addition being made of safflower seed oil, fish meal and vitamine supplement is fed for transgenic positive pig to the F2 of acquisition
The feed of agent；
The transgene pig is to turn fat-1 gene pigs.
2. restorative procedure according to claim 1, which is characterized in that the feed contains based on 100kg：Kardiseed is edible
Oily 3-3.5kg, fish meal 3-3.5kg and vitamine supplement 7-9g.
3. the safflower seed oil compound additive containing safflower seed oil turns fat-1 gene pig gene integrations position in preparation for repairing
Application in the feed of effect side effect, which is characterized in that the safflower seed oil compound additive includes：Edible safflower-seed oil,
Fish meal and vitamine supplement, the feed contain based on 100kg：Edible safflower-seed oil 3-3.5kg, fish meal 3-3.5kg with
And vitamine supplement 7-9g.
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| CN105211636A (en) * | 2015-08-25 | 2016-01-06 | 杭州香保饲料有限公司 | A kind of hybrid process method of grease, albumen, whey powder, feed addictive |
Non-Patent Citations (4)
| Title |
|---|
| Effects of safflower oil supplementation in diet on growth performance and body fatty acid composition of turbot (Psetta maxima);Mehmet Sukru Altundag, et.al.;《Aquacult Int》;20130817;第22卷;第599页表1 * |
| SPF小型猪血液学、血液生化正常参考值、尿常规值测定;詹纯列，等;《中国实验动物学杂志》;20010930;第11卷(第3期);摘要，第135页第1段，第136页第2段，第137页第2.2节 * |
| 实验用五指山小型猪正常生理值测定;靳洪涛，等;《实验动物科学》;20071231;第24卷(第6期);摘要，表5 * |
| 转sFat1基因猪生理生化指标与繁殖性能测定;华文君，等;《安徽农业科学》;20101231;第38卷(第32期);第18235页第1.2节，表1，第18236页"3 讨论" * |
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