根据已发表的人溶菌酶 (human lysozyme,h L YZ) m RNA序列设计引物 ,以人乳腺第 1链 c DNA为模板 ,用 PCR扩增出 1.5 kb h L YZ双链 c DNA,其推导的氨基酸序列与国外发表的从胎盘、巨噬细胞和结肠中克隆的 h L YZ氨基酸序列同源性为 10...根据已发表的人溶菌酶 (human lysozyme,h L YZ) m RNA序列设计引物 ,以人乳腺第 1链 c DNA为模板 ,用 PCR扩增出 1.5 kb h L YZ双链 c DNA,其推导的氨基酸序列与国外发表的从胎盘、巨噬细胞和结肠中克隆的 h L YZ氨基酸序列同源性为 10 0 % ,与从人组织细胞中克隆的 h L YZ仅有 1个氨基酸差异 ,但与从中国人胎盘中克隆的 h L YZ具有6个氨基酸差异。将此 c DNA克隆入乳腺特异表达载体 ,用所获得的基因构件注射哺乳期小鼠 ,经 RT- PCR和微球菌溶解试验证明 ,上述基因构件能有效地驱动目的基因在小鼠乳腺中表达 ,表达量为 6 9.3mg/ L,表达具有较好的组织特异性。这些试验结果表明 ,本研究构建的表达 h L YZ c DNA的基因构件可以用于乳腺生物反应器的研制。展开更多
Five female transgenic mice were produced by microinjection using a construct made up of a 7.3-kb-5′ flanking region and a 2.0-kb coding region of human a-lactalbumin, as well as a 227-bp 3′-flanking region from bov...Five female transgenic mice were produced by microinjection using a construct made up of a 7.3-kb-5′ flanking region and a 2.0-kb coding region of human a-lactalbumin, as well as a 227-bp 3′-flanking region from bovine growth hormone gene. A founder female ex-pressed human a-lactalbumin as much as 0.3 g per liter of its milk, approximately a 3-fold in-crease in the total a-lactalbumin concentration of the transgenic mouse milk. Compared with the normal mice, the expression profile of the ha-Lac transgene in the transgenics is different during the lactation, showing low level in the first 3 days and becoming increased from day 4, then gradually reaching and stabilizing at the highest level from day 13. In addition, the milk yielding volume in the transgenics tended to be higher than in normal mice, suggesting higher concen-trations of a-lactalbumin might boost more milk output.展开更多
文摘根据已发表的人溶菌酶 (human lysozyme,h L YZ) m RNA序列设计引物 ,以人乳腺第 1链 c DNA为模板 ,用 PCR扩增出 1.5 kb h L YZ双链 c DNA,其推导的氨基酸序列与国外发表的从胎盘、巨噬细胞和结肠中克隆的 h L YZ氨基酸序列同源性为 10 0 % ,与从人组织细胞中克隆的 h L YZ仅有 1个氨基酸差异 ,但与从中国人胎盘中克隆的 h L YZ具有6个氨基酸差异。将此 c DNA克隆入乳腺特异表达载体 ,用所获得的基因构件注射哺乳期小鼠 ,经 RT- PCR和微球菌溶解试验证明 ,上述基因构件能有效地驱动目的基因在小鼠乳腺中表达 ,表达量为 6 9.3mg/ L,表达具有较好的组织特异性。这些试验结果表明 ,本研究构建的表达 h L YZ c DNA的基因构件可以用于乳腺生物反应器的研制。
文摘Five female transgenic mice were produced by microinjection using a construct made up of a 7.3-kb-5′ flanking region and a 2.0-kb coding region of human a-lactalbumin, as well as a 227-bp 3′-flanking region from bovine growth hormone gene. A founder female ex-pressed human a-lactalbumin as much as 0.3 g per liter of its milk, approximately a 3-fold in-crease in the total a-lactalbumin concentration of the transgenic mouse milk. Compared with the normal mice, the expression profile of the ha-Lac transgene in the transgenics is different during the lactation, showing low level in the first 3 days and becoming increased from day 4, then gradually reaching and stabilizing at the highest level from day 13. In addition, the milk yielding volume in the transgenics tended to be higher than in normal mice, suggesting higher concen-trations of a-lactalbumin might boost more milk output.
