拉布拉多犬髋关节发育不良和神经类型相关基因SNP位点的检测
发布时间：2019-06-12 08:12
【摘要】：目的:1、检测拉布拉多犬基因组DNA中与髋关节发育不良密切相关的5个基因中的6个SNP位点,以期为拉布拉多犬的选育提供有效的遗传学诊断方法。2、检测拉布拉多犬基因组DNA中与神经类型密切相关的5个基因中的7个SNP位点,以期为拉布拉多犬神经类型的早期鉴定提供分子遗传学鉴定指标。方法:本研究所选用的犬和其信息均由中国导盲犬大连培训基地(以下简称基地)提供;1、经影像学诊断,筛选出典型的6只髋关节发育不良和11只正常拉布拉多犬作为实验对象,采取静脉血,提取血细胞的基因组DNA样本,采用PCR/测序方法对17个样本的5个基因中的6个SNP位点:LAMA2(CFA1:70997779),LRR1(CFA8:29247021),SCR(CFA24:28944481),COL6A3(CFA25:51031100,51040259)和FN1(CFA37:25095511)进行变异型检测,并计算分型所占的百分比,分析正常犬和髋关节发育不良犬的差异趋势。2、依据基地成熟的导盲犬神经类型行为学评估标准,选取典型的安静、活泼、胆小型(弱型)三种神经类型的犬各6只为研究对象,采取静脉血,提取血细胞的基因组DNA样本,采用PCR-RFLP分析法和PCR/测序方法对18个样本5个基因的7个SNP位点:COMT(G482A)、MAOB(T199C)、SLC1A2(T471C)、ABCB1(A985T、T3442C、377-378 ins C)、GNB1L(961-962 ins G)进行变异型检测,并统计分析SNP位点的变异型与神经类型的关联度。结果:1、LAMA2(CFA1:70997779),LRR1(CFA8:29247021),SCR(CFA24:28944481),COL6A3(CFA25:51031100)和FN1(CFA37:25095511)SNP位点的变异型与拉布拉多犬的髋关节发育不良没有显著差异趋势。COL6A3(CFA25:51040259)SNP位点的A/A型与拉布拉多犬的髋关节发育不良存在明显的差异趋势(30%)。2、COMT基因的G482A SNP位点G/G型在安静型犬中所占比例显著高于胆小型(P=0.026),但该两种类型的犬所占比例与活泼型均不存在显著性差异(P0.05);G/A型在胆小型犬中所占比例显著高于活泼型和安静型(P=0.011,P=0.011);A/A型在活泼型犬中所占比例显著高于安静型和胆小型(P=0.008,P=0.008)。MAOB基因的T199C SNP位点在安静、活泼、胆小这三种类型中所占比例均没有显著差异(P0.05)。SLC1A2基因的T471C SNP位点T/T型在活泼型犬中所占比例显著高于安静型和胆小型(P=0.026,P=0.006);T/C型在活泼型犬中所占比例显著低于安静型和胆小型(P=0.023,P=0.011);C/C型在活泼型犬中所占比例显著低于安静型和胆小型(P=0.008,P=0.008)。ABCB1基因的A985T SNP位点T/T型在活泼型犬中所占比例显著高于胆小型(P=0.014),但该两种类型的犬所占比例与安静型均不存在显著性差异(P0.05);T/A型在活泼型犬中所占比例显著低于安静型和胆小型(P=0.005,P=0.002);而A/A型在安静、活泼、胆小这三种类型中所占比例均没有显著差异(P0.05)。ABCB1基因的T3442C SNP位点T/T型在活泼型犬中所占比例显著高于胆小型(P=0.045),但该两种类型的犬所占比例与安静型均不存在显著性差异(P0.05);T/C型在胆小型、安静型、活泼型犬中所占比例均存在显著性差异(P0.05),且胆小型所占比例最高(50%),活泼型所占比例最低(0%);C/C型在安静、活泼、胆小这三种类型中所占比例均没有显著差异(P0.05)。ABCB1基因的377-378 ins C和GNB1L基因的961-962ins G均没有发现显著性差异(P0.05)。结论:1、COL6A3(CFA25:51040259)SNP位点的A/A型可能成为拉布拉多犬髋关节发育不良的实用性遗传学诊断指标,但尚需大样本确认。2、COMT基因的G482A SNP位点的G/A型和A/A型,SLC1A2基因的T471 SNP位点,ABCB1基因的A985T SNP位点的T/A和ABCB1基因的T3442C SNP位点的T/C型有望成为拉布拉多犬神经类型早期鉴定的分子遗传学诊断指标,但尚需大样本确认。
[Abstract]:Objective:1. To detect the 6 SNP sites in 5 genes closely related to the dysplasia of the hip in the Labrador genomic DNA, with a view to providing an effective genetic diagnosis method for the breeding of Labrador. Seven SNP sites in the 5 genes closely related to the type of nerve in the Labrador genomic DNA were detected, with a view to providing molecular genetic identification for the early identification of the Labrador type of nerve type. Methods: The dog and its information selected by the Institute were provided by the Dalian Training Base (hereinafter referred to as the base) of the Chinese guide dog;1. According to the image diagnosis, the typical 6 total hip dysplasia and 11 normal Labrador were selected as experimental subjects, and venous blood was taken. The genomic DNA samples of the blood cells were extracted and six SNP sites in the five genes of the 17 samples were analyzed by a PCR/ sequencing method: LAM2 (CFA1:70997779), LRR1 (CFA8:29247021), SCR (CFA24:28944481), COL6A3 (CFA25:51031100,51040259) and FN1 (CFA37:25095511), and the percentage of the typing was calculated, The differences of dogs in normal and hip dysplasia were analyzed.2. The typical quiet, active and timid (weak) three types of dogs were selected for the purpose of study, and venous blood was taken for each of the three types of dogs of the type of quiet, active and timid (weak type). The genomic DNA samples of blood cells were extracted, and 7 SNP sites of five genes of 18 samples were detected by PCR-RFLP and PCR/ sequencing methods: COMT (G482A), MAOB (T199C), SLC1A2 (T471C), ABCB1 (A985T, T3442C,377-378 ins C), and GNB1L (961-962 ins G). And the incidence degree of the variant and the nerve type of the SNP locus is analyzed. Results:1, LAM2 (CFA1:70997779), LRR1 (CFA8:29247021), SCR (CFA24:28944481), COL6A3 (CFA25:51031100) and FN1 (CFA37:25095511) SNP site did not have a significant difference in the hip dysplasia of the Labrador. There was a significant difference (30%) between the type A/ A of COL6A3 (CFA25:51040259) and the dysplasia of the hip in the Labrador dog (30%). The proportion of the G482A SNP site G/ G of the COMT gene in the quiet dog was significantly higher than that of the cowardly dog (P = 0.026). However, there was no significant difference between the two types of dogs (P0.05); the proportion of the G/ A type in the cowardly dog was significantly higher than that of the active and the quiet type (P = 0.011, P = 0.011); the proportion of the A/ A in the active dog was significantly higher than that of the quiet and timid (P = 0.008, P = 0.008). The T41C SNP site of the MAOB gene has no significant difference in the three types of quiet, active and timid (P0.05). The proportion of the T471C SNP site T/ T of the SLC1A2 gene in the active dog is significantly higher than that of the quiet and timid (P = 0.026, P = 0.006); The proportion of T/ C in the active dog was significantly lower than that of the quiet and timid (P = 0.023, P = 0.011); the proportion of C/ C in the active dog was significantly lower than that of the quiet and timid (P = 0.008, P = 0.008). The proportion of A985T SNP site T/ T in the ABCB1 gene was significantly higher in the active dog (P = 0.014), but there was no significant difference between the two types of dogs (P = 0.014); the proportion of the T/ A type in the active dog was significantly lower than that of the quiet and timid (P = 0.005, P = 0.002); The ratio of type A/ A in the three types of quiet, active and timid (P0.05). The proportion of T/ T in the T/ T type of the ABCB1 gene in the active dog was significantly higher than that of the cowardly type (P = 0.045), but there was no significant difference between the two types of dogs (P0.05). The proportion of T/ C in the timid, quiet and active dogs was significant (P0.05), and the proportion of the timid (50%) and the active type was the lowest (0%), and the C/ C type was quiet and active. There was no significant difference in the three types of the cowardice (P0.05). No significant difference was found between the 377-378 ins C of the ABCB1 gene and 961-962 ins G of the GNB1L gene (P0.05). Conclusion:1. The A/ A type of the SNP site of COL6A3 (CFA25:51040259) may be a practical genetic diagnostic index for the development of the hip joint in the Labrador, but it still needs to be confirmed by large samples. The G/ A type of the G482A SNP site of the COMT gene and the T471 SNP site of the A/ A type and the SLC1A2 gene, The T/ C type of the T3442C SNP site of the A985T SNP site of the ABCB1 gene is expected to be a molecular genetic diagnostic index for the early identification of the Labrador nerve type, but a large sample confirmation is required.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.292
[Abstract]:Objective:1. To detect the 6 SNP sites in 5 genes closely related to the dysplasia of the hip in the Labrador genomic DNA, with a view to providing an effective genetic diagnosis method for the breeding of Labrador. Seven SNP sites in the 5 genes closely related to the type of nerve in the Labrador genomic DNA were detected, with a view to providing molecular genetic identification for the early identification of the Labrador type of nerve type. Methods: The dog and its information selected by the Institute were provided by the Dalian Training Base (hereinafter referred to as the base) of the Chinese guide dog;1. According to the image diagnosis, the typical 6 total hip dysplasia and 11 normal Labrador were selected as experimental subjects, and venous blood was taken. The genomic DNA samples of the blood cells were extracted and six SNP sites in the five genes of the 17 samples were analyzed by a PCR/ sequencing method: LAM2 (CFA1:70997779), LRR1 (CFA8:29247021), SCR (CFA24:28944481), COL6A3 (CFA25:51031100,51040259) and FN1 (CFA37:25095511), and the percentage of the typing was calculated, The differences of dogs in normal and hip dysplasia were analyzed.2. The typical quiet, active and timid (weak) three types of dogs were selected for the purpose of study, and venous blood was taken for each of the three types of dogs of the type of quiet, active and timid (weak type). The genomic DNA samples of blood cells were extracted, and 7 SNP sites of five genes of 18 samples were detected by PCR-RFLP and PCR/ sequencing methods: COMT (G482A), MAOB (T199C), SLC1A2 (T471C), ABCB1 (A985T, T3442C,377-378 ins C), and GNB1L (961-962 ins G). And the incidence degree of the variant and the nerve type of the SNP locus is analyzed. Results:1, LAM2 (CFA1:70997779), LRR1 (CFA8:29247021), SCR (CFA24:28944481), COL6A3 (CFA25:51031100) and FN1 (CFA37:25095511) SNP site did not have a significant difference in the hip dysplasia of the Labrador. There was a significant difference (30%) between the type A/ A of COL6A3 (CFA25:51040259) and the dysplasia of the hip in the Labrador dog (30%). The proportion of the G482A SNP site G/ G of the COMT gene in the quiet dog was significantly higher than that of the cowardly dog (P = 0.026). However, there was no significant difference between the two types of dogs (P0.05); the proportion of the G/ A type in the cowardly dog was significantly higher than that of the active and the quiet type (P = 0.011, P = 0.011); the proportion of the A/ A in the active dog was significantly higher than that of the quiet and timid (P = 0.008, P = 0.008). The T41C SNP site of the MAOB gene has no significant difference in the three types of quiet, active and timid (P0.05). The proportion of the T471C SNP site T/ T of the SLC1A2 gene in the active dog is significantly higher than that of the quiet and timid (P = 0.026, P = 0.006); The proportion of T/ C in the active dog was significantly lower than that of the quiet and timid (P = 0.023, P = 0.011); the proportion of C/ C in the active dog was significantly lower than that of the quiet and timid (P = 0.008, P = 0.008). The proportion of A985T SNP site T/ T in the ABCB1 gene was significantly higher in the active dog (P = 0.014), but there was no significant difference between the two types of dogs (P = 0.014); the proportion of the T/ A type in the active dog was significantly lower than that of the quiet and timid (P = 0.005, P = 0.002); The ratio of type A/ A in the three types of quiet, active and timid (P0.05). The proportion of T/ T in the T/ T type of the ABCB1 gene in the active dog was significantly higher than that of the cowardly type (P = 0.045), but there was no significant difference between the two types of dogs (P0.05). The proportion of T/ C in the timid, quiet and active dogs was significant (P0.05), and the proportion of the timid (50%) and the active type was the lowest (0%), and the C/ C type was quiet and active. There was no significant difference in the three types of the cowardice (P0.05). No significant difference was found between the 377-378 ins C of the ABCB1 gene and 961-962 ins G of the GNB1L gene (P0.05). Conclusion:1. The A/ A type of the SNP site of COL6A3 (CFA25:51040259) may be a practical genetic diagnostic index for the development of the hip joint in the Labrador, but it still needs to be confirmed by large samples. The G/ A type of the G482A SNP site of the COMT gene and the T471 SNP site of the A/ A type and the SLC1A2 gene, The T/ C type of the T3442C SNP site of the A985T SNP site of the ABCB1 gene is expected to be a molecular genetic diagnostic index for the early identification of the Labrador nerve type, but a large sample confirmation is required.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.292
【参考文献】
相关期刊论文 前10条
1 赵明媛;韩芳;李雅婵;俞剑熊;张雅丽;周子娟;王福金;王爱国;王靖宇;;拉布拉多犬髋关节发育不良相关基因的SNP位点的检测[J];实验动物科学;2015年04期
2 赵明媛;俞剑熊;韩芳;李雅婵;张雅丽;周子娟;王福金;王爱国;王靖宇;;拉布拉多犬神经类型相关基因SNP的分析[J];实验动物科学;2015年03期
3 俞剑熊;张雅丽;周子娟;赵明媛;韩芳;王福金;王爱国;王靖宇;;犬毛色、毛色基因型及性别与导盲犬培训成功率的相关性研究[J];实验动物科学;2014年06期
4 俞剑熊;张雅丽;周子娟;王福金;王爱国;王靖宇;;拉布拉多犬与金毛猎犬毛色基因MC1R(R306ter)与TYRP1(Q331ter)SNP位点的检测[J];实验动物科学;2014年03期
5 王福金;王爱国;王亮;李慧玲;董建一;王靖宇;;偏侧性测试在预测成功导盲犬中的应用[J];实验动物科学;2012年03期
6 王爱国;王福金;王亮;李慧玲;董建一;王靖宇;;气质测试在导盲犬培育中的应用[J];畜牧与兽医;2012年02期
7 马昆;张娈松;卢德章;范宏刚;马海濵;王洪斌;;犬髋关节发育不良的诊断、遗传控制和预防管理[J];黑龙江畜牧兽医;2011年03期
8 冯烁;沈向真;;犬髋关节发育异常的研究进展[J];中国畜牧兽医;2010年08期
9 杜晓鹏;顾_g;;赏析拉布拉多犬与金毛猎犬[J];中国工作犬业;2010年07期
10 翟，
本文编号：2497866
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2497866.html
最近更新
教材专著
