Xue G P, Patel M, Johnson J S, Smyth D J, Vickers C E
CSIRO Plant Industry, 120 Meiers Road, Indooroopilly, Brisbane, Australia.
Plant Cell Rep. 2003 Jul;21(11):1088-94. doi: 10.1007/s00299-003-0627-4. Epub 2003 Apr 16.
The use of barley grains as bioreactors for high-level production of cellulase (1,4-beta-glucanase) was investigated. A hybrid cellulase gene, cel-hyb1, driven by the rice GluB-1 promoter was expressed specifically in developing endosperm. Codon usage optimisation of cel-hyb1 increased its expression in barley grains 527-fold and led to cellulase production of up to 1.5% of total grain protein. CEL-HYB1 enzyme in barley grains was highly stable during post-harvest storage. Selectable marker gene ( hph) was subsequently eliminated from transgenic lines through segregation of hph from synthetic cel-hyb1 ( syn.cel-hyb1) in T1 progeny, using a binary plasmid containing hph and syn.cel-hyb1 in separate T-DNAs. These data suggest that barley grains can potentially be used for the commercial production of cellulase.
研究了利用大麦籽粒作为生物反应器高效生产纤维素酶（1,4-β-葡聚糖酶）的情况。由水稻GluB-1启动子驱动的杂种纤维素酶基因cel-hyb1在发育中的胚乳中特异性表达。对cel-hyb1进行密码子使用优化后，其在大麦籽粒中的表达提高了527倍，纤维素酶产量高达总籽粒蛋白的1.5%。大麦籽粒中的CEL-HYB1酶在收获后储存期间高度稳定。随后，使用在单独的T-DNA中含有hph和合成cel-hyb1（syn.cel-hyb1）的二元质粒，通过T1代子代中hph与syn.cel-hyb1的分离，从转基因品系中消除了选择标记基因（hph）。这些数据表明大麦籽粒有潜力用于纤维素酶的商业生产。
