橡胶树半胱氨酸蛋白酶基因HbCP2的克隆与表达特性分析

Molecular Cloning and Expression Analysis of a Cysteine Proteinase Gene (HbCP2) from Hevea brasiliensis

  • 摘要: 根据EST和基因组序列设计引物,应用RT-PCR技术从橡胶树的根组织中分离到1条长1056bp的cDNA,该序列包含了1023bp的开放读码框(ORF),17bp的5’ UTR和16bp的3’ UTR;ORF预测编码340个氨基酸,理论分子量为3752kD,等电点为525,属于液泡定位的分泌型蛋白;蛋白含有1个Inhibitor_I29和1个Peptidase_C1A结构域,隶属于木瓜蛋白酶C1家族;蛋白与蓖麻、杨树中同源蛋白的相似性均在80%以上,将基因命名为HbCP2。表达分析显示,在所分析的根、树皮、木质部、芽、叶片、叶柄和乳管等组织中,HbCP2仅在根中被检测到,表现出明显的组织特异性。

     

    Abstract: Based on the EST and genome sequences, a 1056bp cDNA was isolated from the root tissue of Hevea brasiliensis with specific primers via RTPCR. The sequence contains a 1023bp long open reading frame (ORF), 17 bp 5′UTR and 16 bp 3′UTR. The ORF was predicted to encode 340 amino acids with a theoretical molecular weight (Mw) of 3752 kD and isolectric point (pI) of 5.25; The sequence was predicted to be a secretory protein located to vacuoles; the protein contains one Inhibitor_I29 domain and one Peptidase_C1A domain and can be categoried as the papain C1 family; the protein shares a similarity of more than 80% with the homologues in castor bean (Ricinus communis), poplar (Populus trichocarpa), and then this gene was designated as HbCP2. Semiquantitative RTPCR analysis showed that HbCP2 exhibits a rootspecific expression pattern in examed tissues, i.e. root, bark, xylem, bud, leaf, petiole and laticifer.

     

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