摘要：通過(guò)分子生物學(xué)技術(shù)，測(cè)定豬背長(zhǎng)肌中2個(gè)目的基因即維生素E受體基因PXR和共軛亞油酸受體基因PPARγ的mRNA表達(dá)水平，分析其相對(duì)表達(dá)量與豬肉品質(zhì)的相關(guān)性。 方法 根據(jù)GenBank中公布的已知PXR、PPARγ基因序列，用Primer Premier 5.0、BLAST等軟件設(shè)計(jì)PCR引物，并與參考的引物進(jìn)行比對(duì)，選擇最優(yōu)引物對(duì)。引物經(jīng)合成后用RT-PCR進(jìn)行擴(kuò)增和引物驗(yàn)證。 結(jié)果 在所設(shè)計(jì)的引物中PPARγ引物的擴(kuò)增效果較好，PXR沒(méi)有擴(kuò)增出條帶，故最終PXR選用參考引物。 結(jié)論 通過(guò)探討和優(yōu)化PCR體系中退火溫度、MgCl2 濃度和循環(huán)數(shù)等參數(shù)，建立PXR、PPARγ目的基因的mRNA表達(dá)的半定量RT-PCR體系：PXR最佳擴(kuò)增條件為退火溫度58℃、鎂離子濃度2.5mmol、循環(huán)數(shù)34；PPARγ最佳擴(kuò)增條件是退火溫度59℃、鎂離子濃度2.5mmol、循環(huán)數(shù)31。

關(guān)鍵詞：豬背最長(zhǎng)肌，PXR，PPARγ， RT-PCR

Abstract:Determine the gene expression of PXR and PPARγ in porcine m.longissimus dorsi by Molecular Biology and analysis the relationship between the expression and porcine tenderness. Methods  Based on the known PXR and PPARγ gene sequence reported in GenBank, the primers of which were designed by software such as Primer Premier 5.0, BLAST, etc. Then choice the better one compared with referenced primers. After that, the primers were used to amplified by RT-PCR and could be tested by the experiment. Results  Amplified by RT-PCR, PPARγ’ result was better than PXR’s. So the primer of PPARγ was identified to be designed successfully, and we choice the referenced primer of PXR as what we want. Conclusions Set up the better appropriate conditions of PXR and PPARγ in the semi-quantitative RT- PCR system, such as TM,[Mg2+],cycles and so on. The appropriate conditions in the semi-quantitative RT- PCR system of PXR is 58℃,2.5mmol and 34 cycles; and the appropriate conditions of PPARγ is 59℃,2.5mmol and 31cycles. 

Key words： PXR, PPARγ, RT-PCR, longissimus dorsi

　　　本實(shí)驗(yàn)通過(guò)Trizol法提取豬背最長(zhǎng)肌中總RNA，并對(duì)提取出來(lái)的RNA檢測(cè)后進(jìn)行逆轉(zhuǎn)錄，然后將2個(gè)目的基因PXR和PPARγ進(jìn)行PCR擴(kuò)增，并對(duì)其擴(kuò)增的三個(gè)主要影響因素（即退火溫度、[Mg2+]和循環(huán)數(shù)）進(jìn)行單因素試驗(yàn)，在單因素試驗(yàn)的基礎(chǔ)上進(jìn)一步進(jìn)行正交試驗(yàn)，從而得到優(yōu)化的擴(kuò)增條件。