畢業論文

打賞
當前位置: 畢業論文 > 生物論文 >

5個轉灰飛虱CYP基因果蠅品系的建立和定量PCR驗證

時間:2019-06-30 07:41來源:畢業論文
標基因的實驗組果蠅和未啟動目標基因過量表達的對照組果蠅中灰飛虱CYP6CS2v1、CYP6BD10v2、CYP439A1v3、CYP6FJ1v2和CYP4C72基因的表達量

摘要:將以下五種已構建成功的轉基因載體pUAST-attB-CYP6CS2v1、pUAST-attB-CYP6BD10v2、pUAST-attB-CYP439A1v3、pUAST-attB-CYP6FJ1v2和pUAST-attB-CYP4C72利用顯微注射的方法將其分別導入果蠅胚胎,通過篩選和純化獲得純合的轉基因果蠅品系UAS-CYP6CS2v1、UAS-CYP6BD10v2、UAS-CYP439A1v3、UAS-CYP6FJ1v2和UAS-CYP4C72。通過GAL4/UAS系統,將帶有β-tubulin強啟動子的tub-gal4果蠅品系分別與UAS-CYP6CS2v1、UAS-CYP6BD10v2、UAS-CYP439A1v3、UAS-CYP6FJ1v2和UAS-CYP4C72雜交,獲得全身超量表達灰飛虱CYP6CS2v1、CYP6BD10v2、CYP439A1v3、CYP6FJ1v2和CYP4C72基因的果蠅品系tub>CYP6CS2v1、tub>CYP439A1v3、tub>CYP6FJ1v2、tub>CYP4C72和tub>CYP6BD10v2。利用實時熒光定量PCR的方法,檢測了超量表達目標基因的實驗組果蠅和未啟動目標基因過量表達的對照組果蠅中灰飛虱CYP6CS2v1、CYP6BD10v2、CYP439A1v3、CYP6FJ1v2和CYP4C72基因的表達量。結果顯示:實驗組目標基因CYP6CS2v1、CYP6BD10v2、CYP439A1v3、CYP6FJ1v2和CYP4C72的表達量分別是對照組表達量的9.95、30.39、19.10、33.30和38.42倍,與目標基因序列相似的果蠅內源基因在實驗組和對照組中的表達量顯著低于目標基因的表達量,同時無顯著性差異(P>0.05)。因此,灰飛虱CYP6CS2v1、CYP6BD10v2、CYP439A1v3、CYP6FJ1v2和CYP4C72基因在果蠅中得到了超量表達。36747
關鍵詞:灰飛虱;細胞色素P450基因;轉基因果蠅;定量PCR
Construction of 5 transgenic Drosophila strains with 5 CYP genes from Laodelphax striatellus and Q-PCR verification
Abstract:Constructed transgenic vectors of pUAST-attB-CYP6CS2v1, pUAST-attB-CYP6BD10v2, pUAST-attB-CYP439A1v3, pUAST-attB-CYP6FJ1v2 and pUAST-attB-CYP4C72, were introduced into Drosophila embryos respectively, by microinjection. Homozygous transgenic Drosophila strains of UAS-CYP6CS2v1, UAS-CYP6BD10v2, UAS-CYP439A1v3, UAS-CYP6FJ1v2 and UAS-CYP4C72 were obtained by screening and purification. The tub-gal4 Drosophila melanogaster strain with β-tubulin promoter was genetic crossed with UAS-CYP6CS2v1, UAS-CYP6BD10v2, UAS-CYP439A1v3, UAS-CYP6FJ1v2 and UAS-CYP4C72 respectively to obtain the systemic overexpression strains of tub>CYP6CS2v1, tub>CYP439A1v3, tub>CYP6FJ1v2, tub>CYP4C72 and tub>CYP6BD10v2. By using the real-time quantitative PCR method, the expression levels of target genes in the experimental groups were detected. The results showed that the expression levels of CYP6CS2v1, CYP6BD10v2, CYP439A1v3,CYP6FJ1v2 and CYP4C72  in the experimental groups were 9.95, 30.39, 19.10, 33.30 and 38.42 times than that in control groups. The expression levels of endogenous related CYP genes in Drosophila were significantly lower than the expression level of target genes. At the same time, there was no significant difference (P > 0.05). Therefore, the CYP6CS2v1, CYP6BD10v2, CYP439A1v3, CYP6FJ1v2 and CYP4C72 genes of small brown planthopper were overexpressed in the Drosophila. 源¥自%六^^維*論-文+網=www.aftnzs.live
Key words: Laodelphax striatellus; cytochrome P450 ; transgenic Drosophila;Q-PCR
目  錄
摘要    1
關鍵詞    1
Abstract    1
Key words    1
引言    1
材料與方法    2
1.1 供試果蠅品系及飼養方法    2
1.1.1  供試果蠅品系    2
1.1.2  果蠅的飼養    2
1.2 主要試劑與儀器    3
1.2.1 實驗主要試劑    3
1.2.1 實驗儀器設備    3
1.3顯微注射    3
1.3.1制作產卵碟    3
1.3.2 新鮮胚胎的收集    3
1.3.3顯微注射    4
1.4轉基因果蠅品系的純化    4
1.4.1與W1118果蠅雜交    4
1.4.2與平衡子雜交    4 5個轉灰飛虱CYP基因果蠅品系的建立和定量PCR驗證:http://www.aftnzs.live/shengwu/20190630/35318.html
------分隔線----------------------------
推薦內容
双色球走势图带连线