本文選題：豬應激綜合癥 + 高分辨率熔解曲線�。� 參考：《山西農業(yè)大學》2015年碩士論文

【摘要】：目的：氟烷基因(Hal)是導致豬發(fā)生應激綜合癥(porcine stress syndrome, PSS)的一個主效基因,豬應激綜合癥能夠直接影響到豬的肉質,應激敏感的個體在運輸過程、驅趕甚至配種等應激因子的作用下會突發(fā)惡性高熱癥甚至導致死亡,另外,應激敏感個體豬在經過屠宰后會生成白肌肉,導致肉質變差,從而給養(yǎng)豬戶帶來一定的經濟損失。本實驗的目的,就是建立一種簡易、快捷、準確、自動化程度高的檢測Hal的方法。方法：本實驗首先根據GenBank中PSS RYR1基因序列,結合突變位點,合成C/C型和T/T型兩種標準質粒,將C/C、T/T型質粒等比例混勻后,即可得C/T型質粒。然后以構建的質粒為標準,建立HRM小片段和非標記性探針法兩種檢測Hal的高分辨率熔解曲線(High Resolution Melting, HRM)方法。用建立的兩種方法對300頭豬進行檢測。為了驗證這兩種檢測方法的準確性,用目前最常用的PCR-RFLP方法也對這300頭豬進行了檢測。結果：①成功的構建了三種標準質粒②成功的構建了HRM小片段法,檢測結果顯示300份樣品中均沒有氟烷基因的攜帶者。③成功的構建了HRM非標記性探針法,檢測結果顯示300份樣品中沒有氟烷基因的攜帶者。④PCR-RFLP檢測結果顯示300份樣品中沒有氟烷基因的攜帶者。結論：三種方法的檢測結果一致,即用高分辨率熔解曲線檢測Hal是可行的。
[Abstract]:Objective: halothane gene is a major gene responsible for porcine stress syndrome (PSSs) in pigs. Porcine stress syndrome can directly affect the meat quality of pigs, and stress-sensitive individuals are involved in the transport process. Under the action of stress factors, such as driving out and even mating, it may burst into malignant hyperpyrexia or even lead to death. In addition, after slaughtering, stress-sensitive individual pigs will produce white muscles, resulting in poor meat quality, which will bring certain economic losses to pig farmers. The purpose of this experiment is to establish a simple, fast, accurate and automatic method for detecting Hal. Methods: according to the sequence of PSS RYR1 gene in GenBank, two standard plasmids of C / C type and T / T type were synthesized according to the mutation site. The C / T type plasmids were obtained by mixing the C / C / T / T type plasmids. Then, using the constructed plasmids as the standard, two high resolution melting curve (HRM) methods for detecting Hal by HRM small fragment and unlabeled probe method were established. The established two methods were used to detect 300 pigs. In order to verify the accuracy of the two methods, 300 pigs were tested with the most commonly used PCR-RFLP method. Results 1 successfully constructed three standard plasmids 2 and constructed HRM small fragment method successfully. The results showed that 3. 3 carriers without halothane gene in 300 samples successfully constructed HRM unlabeled probe method. The results showed that 300 samples had no Halothane gene carriers. 4 PCR-RFLP analysis showed that there was no Halothane gene carrier in 300 samples. Conclusion: it is feasible to detect Hal by high resolution melting curve.
【學位授予單位】：山西農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.28

【參考文獻】

相關期刊論文 前8條

1 杜曉婷;劉銳;張水明;楊衛(wèi)景;吳正紅;陸建中;金建芳;;PCR-RFLP技術檢測嘉興鳳橋種豬氟烷基因[J];安徽農業(yè)科學;2009年24期

2 王s，

本文編號：1987791