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  本文選題：微型鼠 + ENU誘變�。� 參考：《延邊大學》2006年碩士論文

【摘要】：為進一步研究基因突變與侏儒癥的關系，利用化學物質(zhì)乙基亞硝基脲(N-ethyl-N-nitrosourea，ENU)人工誘變了BALB／c小鼠，經(jīng)傳代產(chǎn)生了一種侏儒癥形態(tài)的小鼠，并命名這種變異小鼠為微型鼠(Small Body Size mouse，SBS)。經(jīng)遺傳分析發(fā)現(xiàn)微型鼠表現(xiàn)常染色體隱性遺傳特性。微型鼠在出生1日齡時，其形態(tài)與正常BALB／c小鼠肉眼不易區(qū)分，但其體重與正常BALB／c小鼠比較差異顯著。1周齡后的微型鼠經(jīng)肉眼觀察其形態(tài)可與正常BALB／c小鼠區(qū)分，并且體重差異更加顯著。本試驗不但檢測了微型鼠的體重及骨密度變化，而且利用微衛(wèi)星遺傳標記通過遺傳連鎖分析對突變基因進行了初步定位。試驗中隨機選擇了30只微型鼠和24只正常BALB／c小鼠，從出生到14周齡進行長期的觀察并記錄體重變化；并且另外隨機選擇了5只微型鼠和5只正常小鼠，分別在3周齡和6周齡進行骨密度檢測。為了實現(xiàn)對突變基因的初步定位，本研究首先利用回交試驗法培育出了142只回交后代[(SBS×C57BL／6J)F_1×SBS]F_2，再次通過PCR法對19條常染色體上的79個微衛(wèi)星遺傳標記進行詳細的篩選，從而完成遺傳連鎖分析。試驗結果表明，，從出生到14周齡微型鼠的體重顯著低于同齡的正常BALB／c小鼠的體重，尤其在3周齡時其體重是正常BALB／c小鼠體重的43.7％，差異極顯著(P＜0.001)。在骨密度檢測的結果中，本研究發(fā)現(xiàn)微型鼠在3周齡和6周齡時的骨密度都明顯低于同齡正常BALB／c小鼠的。遺傳連鎖分析結果表明，突變基因(sbs)初步定位在第10條染色體上，選用本染色體上與侏儒病表型相關基因產(chǎn)后體重增長因子(Pbwg9)和過氧化體生物合成因子(Pex7)最近的微衛(wèi)星D10Mit283標記引物擴增，在142只F_2代中只發(fā)生一例交換，表明Pbwg9基因和Pex7基因是本實驗中侏儒癥突變的強有力的候選基因。
[Abstract]:In order to further study the relationship between gene mutation and dwarfism, BALB/c mice were induced by the chemical substance ethyl-N-nitrosourea- (ENUU), and a dwarfism morphologic mouse was produced by subculture. The mutated mice were named as small Body Size mousetrap (SBSN). Genetic analysis revealed that micromice showed autosomal recessive genetic characteristics. At 1 day of birth, the morphology of miniature mice was not easily distinguished from that of normal BALB/c mice, but their body weight was significantly different from that of normal BALB/c mice. 1 weeks after birth, the morphology of miniature mice could be distinguished from that of normal BALB/c mice by naked eye observation. And the weight difference is even more significant. Not only the body weight and bone mineral density of micromice were detected, but also the mutated genes were preliminarily located by genetic linkage analysis using microsatellite genetic markers. In the experiment, 30 miniature mice and 24 normal BALB/c mice were randomly selected for long-term observation from birth to 14 weeks of age and body weight changes were recorded, and 5 miniature mice and 5 normal mice were randomly selected. Bone mineral density (BMD) was measured at 3 weeks and 6 weeks of age, respectively. In order to locate the mutated genes, we first selected 142 backcross progenies [SBS 脳 C57BL/6J)F_1 脳 SBS] F2 by backcross test, and then screened 79 microsatellite genetic markers on 19 autosomes by PCR method. Thus, genetic linkage analysis was completed. The results showed that the body weight of the miniature mice from birth to 14 weeks was significantly lower than that of the normal BALB/c mice of the same age, especially at the age of 3 weeks, the body weight was 43.7 of the normal BALB/c mice, and the difference was very significant (P < 0.001). The results of bone mineral density (BMD) test showed that the BMD of miniature mice at the age of 3 weeks and 6 weeks was significantly lower than that of the normal BALB/c mice of the same age. The results of genetic linkage analysis showed that the mutant gene was located on chromosome 10. Microsatellite D10Mit283 marker primers were used to amplify the gene associated with dwarf disease phenotypic postpartum weight gain factor (Pbwg9) and peroxisome biosynthesis factor (Pex7) on this chromosome. Only one exchange occurred in 142 F2s. The results showed that Pbwg9 gene and Pex7 gene were strong candidate genes for dwarfism mutation in this experiment.
【學位授予單位】：延邊大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R-332

【參考文獻】

相關期刊論文 前1條

1 吳寶金,茅慧華,李厚達;ENU誘變在功能基因組研究中的應用[J];癌變.畸變.突變;2004年01期

本文編號：1880928