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

【摘要】：為進(jìn)一步研究基因突變與侏儒癥的關(guān)系，利用化學(xué)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(shí)，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(guān)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(yàn)不但檢測(cè)了微型鼠的體重及骨密度變化，而且利用微衛(wèi)星遺傳標(biāo)記通過(guò)遺傳連鎖分析對(duì)突變基因進(jìn)行了初步定位。試驗(yàn)中隨機(jī)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(jìn)行長(zhǎng)期的觀(guān)察并記錄體重變化；并且另外隨機(jī)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(jìn)行骨密度檢測(cè)。為了實(shí)現(xiàn)對(duì)突變基因的初步定位，本研究首先利用回交試驗(yàn)法培育出了142只回交后代[(SBS×C57BL／6J)F_1×SBS]F_2，再次通過(guò)PCR法對(duì)19條常染色體上的79個(gè)微衛(wèi)星遺傳標(biāo)記進(jìn)行詳細(xì)的篩選，從而完成遺傳連鎖分析。試驗(yàn)結(jié)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(shí)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(cè)的結(jié)果中，本研究發(fā)現(xiàn)微型鼠在3周齡和6周齡時(shí)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(jié)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(guān)基因產(chǎn)后體重增長(zhǎng)因子(Pbwg9)和過(guò)氧化體生物合成因子(Pex7)最近的微衛(wèi)星D10Mit283標(biāo)記引物擴(kuò)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(shí)驗(yàn)中侏儒癥突變的強(qiáng)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(xué)位授予單位】：延邊大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R-332

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 吳寶金,茅慧華,李厚達(dá);ENU誘變?cè)诠δ芑蚪M研究中的應(yīng)用[J];癌變.畸變.突變;2004年01期

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