拉布拉多犬髖關(guān)節(jié)發(fā)育不良和神經(jīng)類型相關(guān)基因SNP位點的檢測
發(fā)布時間：2019-06-12 08:12
【摘要】：目的:1、檢測拉布拉多犬基因組DNA中與髖關(guān)節(jié)發(fā)育不良密切相關(guān)的5個基因中的6個SNP位點,以期為拉布拉多犬的選育提供有效的遺傳學診斷方法。2、檢測拉布拉多犬基因組DNA中與神經(jīng)類型密切相關(guān)的5個基因中的7個SNP位點,以期為拉布拉多犬神經(jīng)類型的早期鑒定提供分子遺傳學鑒定指標。方法:本研究所選用的犬和其信息均由中國導盲犬大連培訓基地(以下簡稱基地)提供;1、經(jīng)影像學診斷,篩選出典型的6只髖關(guān)節(jié)發(fā)育不良和11只正常拉布拉多犬作為實驗對象,采取靜脈血,提取血細胞的基因組DNA樣本,采用PCR/測序方法對17個樣本的5個基因中的6個SNP位點:LAMA2(CFA1:70997779),LRR1(CFA8:29247021),SCR(CFA24:28944481),COL6A3(CFA25:51031100,51040259)和FN1(CFA37:25095511)進行變異型檢測,并計算分型所占的百分比,分析正常犬和髖關(guān)節(jié)發(fā)育不良犬的差異趨勢。2、依據(jù)基地成熟的導盲犬神經(jīng)類型行為學評估標準,選取典型的安靜、活潑、膽小型(弱型)三種神經(jīng)類型的犬各6只為研究對象,采取靜脈血,提取血細胞的基因組DNA樣本,采用PCR-RFLP分析法和PCR/測序方法對18個樣本5個基因的7個SNP位點:COMT(G482A)、MAOB(T199C)、SLC1A2(T471C)、ABCB1(A985T、T3442C、377-378 ins C)、GNB1L(961-962 ins G)進行變異型檢測,并統(tǒng)計分析SNP位點的變異型與神經(jīng)類型的關(guān)聯(lián)度。結(jié)果:1、LAMA2(CFA1:70997779),LRR1(CFA8:29247021),SCR(CFA24:28944481),COL6A3(CFA25:51031100)和FN1(CFA37:25095511)SNP位點的變異型與拉布拉多犬的髖關(guān)節(jié)發(fā)育不良沒有顯著差異趨勢。COL6A3(CFA25:51040259)SNP位點的A/A型與拉布拉多犬的髖關(guān)節(jié)發(fā)育不良存在明顯的差異趨勢(30%)。2、COMT基因的G482A SNP位點G/G型在安靜型犬中所占比例顯著高于膽小型(P=0.026),但該兩種類型的犬所占比例與活潑型均不存在顯著性差異(P0.05);G/A型在膽小型犬中所占比例顯著高于活潑型和安靜型(P=0.011,P=0.011);A/A型在活潑型犬中所占比例顯著高于安靜型和膽小型(P=0.008,P=0.008)。MAOB基因的T199C SNP位點在安靜、活潑、膽小這三種類型中所占比例均沒有顯著差異(P0.05)。SLC1A2基因的T471C SNP位點T/T型在活潑型犬中所占比例顯著高于安靜型和膽小型(P=0.026,P=0.006);T/C型在活潑型犬中所占比例顯著低于安靜型和膽小型(P=0.023,P=0.011);C/C型在活潑型犬中所占比例顯著低于安靜型和膽小型(P=0.008,P=0.008)。ABCB1基因的A985T SNP位點T/T型在活潑型犬中所占比例顯著高于膽小型(P=0.014),但該兩種類型的犬所占比例與安靜型均不存在顯著性差異(P0.05);T/A型在活潑型犬中所占比例顯著低于安靜型和膽小型(P=0.005,P=0.002);而A/A型在安靜、活潑、膽小這三種類型中所占比例均沒有顯著差異(P0.05)。ABCB1基因的T3442C SNP位點T/T型在活潑型犬中所占比例顯著高于膽小型(P=0.045),但該兩種類型的犬所占比例與安靜型均不存在顯著性差異(P0.05);T/C型在膽小型、安靜型、活潑型犬中所占比例均存在顯著性差異(P0.05),且膽小型所占比例最高(50%),活潑型所占比例最低(0%);C/C型在安靜、活潑、膽小這三種類型中所占比例均沒有顯著差異(P0.05)。ABCB1基因的377-378 ins C和GNB1L基因的961-962ins G均沒有發(fā)現(xiàn)顯著性差異(P0.05)。結(jié)論:1、COL6A3(CFA25:51040259)SNP位點的A/A型可能成為拉布拉多犬髖關(guān)節(jié)發(fā)育不良的實用性遺傳學診斷指標,但尚需大樣本確認。2、COMT基因的G482A SNP位點的G/A型和A/A型,SLC1A2基因的T471 SNP位點,ABCB1基因的A985T SNP位點的T/A和ABCB1基因的T3442C SNP位點的T/C型有望成為拉布拉多犬神經(jīng)類型早期鑒定的分子遺傳學診斷指標,但尚需大樣本確認。
[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.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：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.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S858.292
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