發(fā)布時間：2018-03-09 00:14

  本文選題：山羊　切入點：Agouti基因　出處：《河北農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：本試驗以云南黑山羊的Agouti基因序列為模板設(shè)計引物,利用黑白兩種顏色的山羊基因組作為樣品對Agouti基因部分序列進行擴增,獲得了Agouti基因的部分序列。根據(jù)不同毛色山羊Agouti基因5′非翻譯區(qū)外顯子1剪接體不同類型,設(shè)計引物,采用黑白兩種顏色的山羊個體進行剪接體驗證。試驗結(jié)果表明:所選山羊個體的剪接體類型均為It It′。山羊基因組序列(登錄號:AJPT01136617.1)中有兩段序列與牛的預(yù)測啟動子同源性比較高,因此設(shè)計引物擴增目的片段A和B,同時構(gòu)建兩個熒光素酶報告基因質(zhì)粒PGL3-basic-A和PGL3-basic-B。并用構(gòu)建的熒光素酶報告基因質(zhì)粒瞬時轉(zhuǎn)染人皮膚黑素瘤細胞A375(皮膚細胞)和293T細胞(非皮膚細胞)來檢測兩段目的片段是否具有Agouti基因啟動子活性。結(jié)果用SPSS19.0軟件進行分析,結(jié)果表明:所構(gòu)建的目的片段的質(zhì)粒與陰性對照相比不存在顯著性差異,說明所獲得的兩段目的片段均不具有啟動子活性。
[Abstract]:In this experiment, the Agouti gene sequence of Yunnan black goat was used as template to design primers, and the partial sequence of Agouti gene was amplified by using black and white goat genome as samples. Partial sequence of Agouti gene was obtained. Primers were designed according to different types of splicing of 5 'untranslated region exon 1 of Agouti gene in different wool goats. The experimental results showed that the splicing types of goat individuals were all it It.Goat genome sequence (accession No.: AJPT01136617.1) had high homology with the predicted promoter of cattle. Therefore, primers were designed to amplify the target fragments A and B, and two luciferase reporter gene plasmids PGL3-basic-A and PGL3-basic-Bwere constructed at the same time. The constructed plasmids of luciferase reporter gene were transiently transfected into human skin melanoma cells A375 (skin cells) and 293T thin cells. Cells (non-skin cells) were used to detect whether the two target fragments had the promoter activity of Agouti gene. The results were analyzed by SPSS19.0 software. The results showed that there was no significant difference between the plasmid of the constructed target fragment and that of the negative control, indicating that neither of the two fragments had promoter activity.
【學(xué)位授予單位】：河北農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S827

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本文編號：1586161