Yang Suk-Hwan, Moran Daniel L, Jia Hong-Wu, Bicar Earl H, Lee Michael, Scott M Paul
Interdepartmental Genetics Program, Iowa State University, USA.
Transgenic Res. 2002 Feb;11(1):11-20. doi: 10.1023/a:1013996129125.
The main nutritional limitation of maize used for feed is the content of protein that is digestible, bioavailable and contains an amino acid balance that matches the requirements of animals. In contrast, milk protein has good digestibility, bioavailability and amino acid balance. As an initial effort to create maize optimized as a source of swine nutrition, a codon-adjusted version of a gene encoding the milk protein porcine alpha-lactalbumin was synthesized. Maize expression vectors containing this gene under the control of the Ubi-1 promoter and nos 3' terminator were constructed. These vectors were used to transform maize callus lines that were regenerated into fertile plants. The alpha-lactalbumin transgenes were transmitted through meiosis to the sexual progeny of the regenerated plants. Porcine alpha-lactalbumin was detected in callus and kernels from transgenic maize lines that were transformed by two constructs containing the 27-kDa maize gamma-zein signal sequence at the 5' end of the synthetic porcine alpha-lactalbumin coding sequence. One of these constructs contained an ER retention signal and the other did not. Expression was not observed in kernels or callus from transgenic maize lines that were transformed by a construct that does not contain an exogenous protein-targeting signal. This suggests that the signal peptide might play an important role in porcine alpha-lactalbumin accumulation in transgenic maize kernels.
用作饲料的玉米的主要营养限制在于其可消化、具有生物利用性且氨基酸平衡符合动物需求的蛋白质含量。相比之下,乳蛋白具有良好的消化率、生物利用性和氨基酸平衡。作为将玉米优化为猪营养来源的初步尝试,合成了编码乳蛋白猪α-乳白蛋白的密码子优化版基因。构建了在Ubi-1启动子和nos 3'终止子控制下包含该基因的玉米表达载体。这些载体用于转化玉米愈伤组织系,后者再生为可育植株。α-乳白蛋白转基因通过减数分裂传递给再生植株的有性后代。在由两种在合成猪α-乳白蛋白编码序列5'端包含27-kDa玉米γ-醇溶蛋白信号序列的构建体转化的转基因玉米系的愈伤组织和籽粒中检测到了猪α-乳白蛋白。其中一种构建体包含内质网滞留信号,另一种则没有。在由不含外源蛋白靶向信号的构建体转化的转基因玉米系的籽粒或愈伤组织中未观察到表达。这表明信号肽可能在转基因玉米籽粒中猪α-乳白蛋白的积累中起重要作用。