Xiao Dong, Yue Ying, Deng Xin-Yan, Huang Bing, Guo Zhong-Min, Ma Yun, Lin Yi-Li, Hong Xun, Tang Huan, Xu Kang, Chen Xi-Gu
Center of Experimental Animals, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China.
World J Gastroenterol. 2007 Jan 14;13(2):244-9. doi: 10.3748/wjg.v13.i2.244.
To use the tyrosinase minigene as a visual marker to perform microinjection training and improve the techniques related with transgene to greatly elevate the efficiency of gene transfer.
A mouse tyrosinase minigene, i.e., TyBS, in which the 2.25-kb authentic genomic 5' non-coding flanking sequence of mouse tyrosinase was fused to a mouse tyrosinase cDNA, was introduced into the fertilized eggs of outbred Kunming albino mice.
Of the 11 animals that developed from the injected eggs, two mice (P1 and #8) exhibited pigmented hair (P1) and eyes (P1 and #8), as confirmed by PCR analysis for the tyrosinase minigene integrated into the genome. When founder P1 was bred to Kunming male mouse, six progeny out of 11 offspring inherited the transgene and the pigmented-eye phenotype.
Taken together, these results suggest that this minigene encodes the active tyrosinase protein and that its 5' flanking region contains the sequences regulating the expression of mouse tyrosinase gene as expected. We have rescued the albino phenotype by introduction and expression of a functional tyrosinase minigene in the Kunming albino mouse and the transgene can be passed to subsequent generation. These findings also indicate that TyBS can be a useful visual marker gene in the co-transgenic experiments.
使用酪氨酸酶小基因作为视觉标记进行显微注射训练，并改进与转基因相关的技术，以大幅提高基因转移效率。
将一个小鼠酪氨酸酶小基因，即TyBS，导入远交群昆明白化小鼠的受精卵中。TyBS是将小鼠酪氨酸酶2.25 kb真实基因组5'非编码侧翼序列与小鼠酪氨酸酶cDNA融合而成。
在由注射后的卵发育而来的11只动物中，两只小鼠（P1和#8）表现出有色毛发（P1）和眼睛（P1和#8），通过对整合到基因组中的酪氨酸酶小基因进行PCR分析得以证实。当奠基者P1与昆明雄性小鼠交配时，11只后代中有6只继承了转基因和有色眼表型。
综上所述，这些结果表明该小基因编码活性酪氨酸酶蛋白，并且其5'侧翼区域如预期包含调节小鼠酪氨酸酶基因表达的序列。我们通过在昆明白化小鼠中导入并表达功能性酪氨酸酶小基因挽救了白化病表型，并且转基因可以传递给后代。这些发现还表明TyBS在共转基因实验中可以是一个有用的视觉标记基因。
