本文選題：豬應(yīng)激綜合癥 + 高分辨率熔解曲線��； 參考：《山西農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：目的：氟烷基因(Hal)是導(dǎo)致豬發(fā)生應(yīng)激綜合癥(porcine stress syndrome, PSS)的一個(gè)主效基因,豬應(yīng)激綜合癥能夠直接影響到豬的肉質(zhì),應(yīng)激敏感的個(gè)體在運(yùn)輸過(guò)程、驅(qū)趕甚至配種等應(yīng)激因子的作用下會(huì)突發(fā)惡性高熱癥甚至導(dǎo)致死亡,另外,應(yīng)激敏感個(gè)體豬在經(jīng)過(guò)屠宰后會(huì)生成白肌肉,導(dǎo)致肉質(zhì)變差,從而給養(yǎng)豬戶帶來(lái)一定的經(jīng)濟(jì)損失。本實(shí)驗(yàn)的目的,就是建立一種簡(jiǎn)易、快捷、準(zhǔn)確、自動(dòng)化程度高的檢測(cè)Hal的方法。方法：本實(shí)驗(yàn)首先根據(jù)GenBank中PSS RYR1基因序列,結(jié)合突變位點(diǎn),合成C/C型和T/T型兩種標(biāo)準(zhǔn)質(zhì)粒,將C/C、T/T型質(zhì)粒等比例混勻后,即可得C/T型質(zhì)粒。然后以構(gòu)建的質(zhì)粒為標(biāo)準(zhǔn),建立HRM小片段和非標(biāo)記性探針?lè)▋煞N檢測(cè)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(duì)300頭豬進(jìn)行檢測(cè)。為了驗(yàn)證這兩種檢測(cè)方法的準(zhǔn)確性,用目前最常用的PCR-RFLP方法也對(duì)這300頭豬進(jìn)行了檢測(cè)。結(jié)果：①成功的構(gòu)建了三種標(biāo)準(zhǔn)質(zhì)粒②成功的構(gòu)建了HRM小片段法,檢測(cè)結(jié)果顯示300份樣品中均沒(méi)有氟烷基因的攜帶者。③成功的構(gòu)建了HRM非標(biāo)記性探針?lè)?檢測(cè)結(jié)果顯示300份樣品中沒(méi)有氟烷基因的攜帶者。④PCR-RFLP檢測(cè)結(jié)果顯示300份樣品中沒(méi)有氟烷基因的攜帶者。結(jié)論：三種方法的檢測(cè)結(jié)果一致,即用高分辨率熔解曲線檢測(cè)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(xué)位授予單位】：山西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.28

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相關(guān)期刊論文 前8條

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本文編號(hào)：1987791