Ji Shengjian, Lu Yingchun, Li Jun, Wei Gang, Liang Xiujuan, Zhu Yuxian
State Key Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, 100871, Beijing, China.
Biochem Biophys Res Commun. 2002 Sep 6;296(5):1245-50. doi: 10.1016/s0006-291x(02)02069-7.
Using cDNA Representational Difference Analysis (RDA) techniques, we isolated a cDNA that was expressed specifically in cotton fibers but not in the ovules of a fuzzless-lintless mutant (fl). We designated it as Gh-BTubL for it shares high sequence identity with known plant and yeast beta-tubulins. RT-PCR and robotic cDNA dot blot analyses indicated that the expression of Gh-BTubL was correlated with the elongation pattern of cotton fibers. In situ hybridization results verified that there was no Gh-BTubL mRNA in fl ovules while it was easily detected in the elongating wild type cotton fiber cells. Overexpression of Gh-BTubL in fission yeast induced longitudinal growth of the host cells by 1.74-fold, with no apparent effect on other aspects of the host cells. We suggest that Gh-BTubL plays an important role in cotton fiber elongation and we believe that elucidation of the control mechanisms for expression of tubulin-like proteins may help improve fiber quality and productivity.
利用cDNA代表性差异分析(RDA)技术,我们分离出一个在棉纤维中特异性表达而在无绒无絮突变体(fl)胚珠中不表达的cDNA。我们将其命名为Gh-BTubL,因为它与已知的植物和酵母β-微管蛋白具有高度的序列同一性。RT-PCR和机器人cDNA斑点印迹分析表明,Gh-BTubL的表达与棉纤维的伸长模式相关。原位杂交结果证实,fl胚珠中没有Gh-BTubL mRNA,而在伸长的野生型棉纤维细胞中很容易检测到。在裂殖酵母中过表达Gh-BTubL可使宿主细胞的纵向生长增加1.74倍,对宿主细胞的其他方面没有明显影响。我们认为Gh-BTubL在棉纤维伸长中起重要作用,并且我们相信阐明类微管蛋白样蛋白的表达调控机制可能有助于提高纤维质量和产量。