摘要
采用巴西橡胶树常割树胶乳的Poly(A+)RNA为Tester,叶片Poly(A+)RNA为Driver,通过抑制消减杂交法(suppressivesubtractivehybridization,SSH)构建了一个胶乳特异表达基因差减文库,通过菌落PCR及反式NorthernDot-Blot筛选鉴定阳性差异片段,获得79个胶乳特异表达阳性克隆,部分阳性克隆还通过NorthernBlot杂交及RT-PCR进一步验证。随机挑选部分阳性克隆序列比较分析,结果表明有部分克隆与巴西橡胶树中已知的基因相同,而多数克隆在巴西橡胶树中是新发现,它们与橡胶生物合成、物质代谢运输、信号传导和形态建成等相关。
A subtracted cDNA library was constructed via suppressive subtractive hybridization（SSH）, in which the latex Poly （A ＋ ）RNA isolated from regularly tapped Hevea brasiliensis trees was utilized as tester while driver is Poly（A ＋ ）RNA derived from negative latex-producing leaf tissues. Total 79 positive clones were identified from screening of the subtracted library by PCR and Reverse Northern Dot-Blot, part of them were further verified by RT-PCR and Northern Blot. 15 out of these 79 confirmed clones were randomly selected for sequencing analyses, of which the results were aligned with GenBank databases via BLASTN and BLASTX programs. It is shown that 5 of 15 candidates were identity with known genes in H. brasiliensis which exerted a positive role in rubber synthesis, 2 showed significant similarity to cis-prenyltransferase of H. brasiliensis, 11 were novel genes in H. brasiliensis.
出处
《林业科学》
EI
CAS
CSCD
北大核心
2006年第6期32-36,共5页
Scientia Silvae Sinicae
基金
中国热带农业科学院科技基金项目(Rykj0420)。
