Jefferson R A, Klass M, Wolf N, Hirsh D
J Mol Biol. 1987 Jan 5;193(1):41-6. doi: 10.1016/0022-2836(87)90624-3.
We have shown the expression of transformed genes in the nematode Caenorhabditis elegans using a new gene fusion system. Vectors consisting of the flanking regions of a collagen gene (col-1) or a major sperm protein gene of C. elegans fused to the Escherichia coli uidA gene, encoding beta-glucuronidase, were microinjected into worms and found to be propagated as high-copy extrachromosomal tandem arrays. We have detected beta-glucuronidase activity in transformed lines, and have shown that the activity is dependent upon the correct reading frame of the construction and on the presence of the worm sequences. The enzyme activity was shown to be encoded by the chimeric beta-glucuronidase gene by co-segregation analysis and by inactivation with specific antisera. Expression is at a very low level, and seems to be constitutive. We have used histochemical techniques to visualize the enzyme activity in embryos.
我们利用一种新的基因融合系统,展示了转化基因在秀丽隐杆线虫中的表达情况。将由秀丽隐杆线虫胶原蛋白基因(col-1)或主要精子蛋白基因的侧翼区域与编码β-葡萄糖醛酸酶的大肠杆菌uidA基因融合而成的载体显微注射到线虫体内,发现其以高拷贝数的染色体外串联阵列形式进行增殖。我们在转化品系中检测到了β-葡萄糖醛酸酶活性,并表明该活性取决于构建体的正确阅读框以及线虫序列的存在。通过共分离分析和用特异性抗血清使其失活,证明了该酶活性由嵌合β-葡萄糖醛酸酶基因编码。表达水平非常低,且似乎是组成型的。我们已经使用组织化学技术来观察胚胎中的酶活性。