从麻疯树胚乳cDNA文库中筛选分离了一个编码酰基载体蛋白的cDNA序列,全长为806bp,其开放读码框(ORF)长度为393bp,编码130个氨基酸,该序列在GenBank登录号为EF179617,命名为JcACP.该基因编码的蛋白质相对分子质量(Mr)约为14.4×10^3,等电点为5.2,具有酰基载体蛋白的典型特征——含有4'-磷酸泛酰巯基乙胺辅基的结合位点.RT-PCR试验结果表明,该基因在麻疯树的叶、茎和种子中表达,以种子中的表达量最高,在根中不表达,种子萌发后其表达量随萌发进程递增,在96h时表达量达到最高.结果表明,JcACP在种子中的高度表达可能与种子萌发时的代谢活动有关.同时,将其在大肠杆菌中成功进行了原核表达.图5参18
A cDNA clone encoding ACP (acyl carrier protein) was isolated from Jatropha curcas L. endosperm cDNA library. The sequence was 806 bp in length and the ORF was 393 bp. It encoded a putative protein of 130 amino acids and was named as JcACP. The predicted molecular weight (M) of the putative protein encoded by JcACP was 14.4×10^3, pi=5.2. It possessed all the typical characteristics of ACPs, containing a 4'- phosphopantetheine-binding motif. RT-PCR analysis showed that JcACP gene was expressed in the leaves, stems and seeds of J. curcas. The expression level of JcACP was the highest in seeds and it was not detected in roots. After germination, the expression level of JcACP in seeds increased progressively and reached a peak at 96 h. These results indicated that the expression of JcACP in the seeds was probably related to the seeds metabolism during the germination. It was expressed in Escherichia coli successfully for further research. Fig 5, Ref 18