摘要
为了探究马尾松木材形成的分子机制,以马尾松嫩枝的总RNA反转录得到的cDNA为模板,利用反转录PCR和RACE技术克隆得到马尾松CesA2基因的全长cDNA序列(命名为Pm Ces A2),序列长度为3 500 bp,包含一个长为3 174 bp的开放阅读框(open reading frame,ORF),编码1 057个氨基酸。序列分析表明:PmCesA2序列与已报道的火炬松Ces A2基因(AY789651.1)的相似性达98%。将全长基因克隆到载体p BI121质粒的Sna BI,Sac I双酶切位点之间,构建正义表达载体p BI121-PmCesA2。
In order to discover the molecular mechanism of wood formation in Pinus massoniana, total RNA extracted from twigs of P. massoniana was reverse transcribed into cDNA and used as PCR template. With the strategies of RT-PCR and RACE, we cloned full-length CesA2 gene cDNA （ named PmCesA2 ） that was 3 500 bp in length with a 3 174 bp open rea- ding frame （ ORF）, encoding 1 057 amino acids. Sequence analysis showed that PmCesA2 presented a 98% similarity with reported CesA2 gene （AY789651.1） of P. taeda. To take advantage of the plasmid pBI121 plant expression vector for the construction, double digested by SnaBI and SacI, PmCesA2 gene nucleotide sequence was subcloned into SnaBI-Sacl site of pBI121 vector in reverse orientation, which resulted in plasmid pBI121-PmCesA2.
出处
《林业科技开发》
北大核心
2015年第3期11-16,共6页
China Forestry Science and Technology
基金
国家"十二五"科技支撑项目(2012BAD01B0202)
江苏高校优势学科建设工程资助项目(PAPD)
国家林业公益性行业科研专项(201104010)
