發(fā)布時(shí)間：2018-03-09 00:14

  本文選題：山羊　切入點(diǎn)：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(yàn)以云南黑山羊的Agouti基因序列為模板設(shè)計(jì)引物,利用黑白兩種顏色的山羊基因組作為樣品對(duì)Agouti基因部分序列進(jìn)行擴(kuò)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(jì)引物,采用黑白兩種顏色的山羊個(gè)體進(jìn)行剪接體驗(yàn)證。試驗(yàn)結(jié)果表明:所選山羊個(gè)體的剪接體類型均為It It′。山羊基因組序列(登錄號(hào):AJPT01136617.1)中有兩段序列與牛的預(yù)測(cè)啟動(dòng)子同源性比較高,因此設(shè)計(jì)引物擴(kuò)增目的片段A和B,同時(shí)構(gòu)建兩個(gè)熒光素酶報(bào)告基因質(zhì)粒PGL3-basic-A和PGL3-basic-B。并用構(gòu)建的熒光素酶報(bào)告基因質(zhì)粒瞬時(shí)轉(zhuǎn)染人皮膚黑素瘤細(xì)胞A375(皮膚細(xì)胞)和293T細(xì)胞(非皮膚細(xì)胞)來檢測(cè)兩段目的片段是否具有Agouti基因啟動(dòng)子活性。結(jié)果用SPSS19.0軟件進(jìn)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(duì)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(dòng)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(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S827

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本文編號(hào)：1586161