DNA methylation of fly genes and transposons

The use of anti-5-methylcytosine antibodies in affinity columns allowed the identification of methylated sequences in the genome of Drosophila melanogaster adults. In view of the presence of transposable elements amongst the identified sequences, it has been suggested that DNA methylation is involved in transposon control in the fly genome. On the contrary, a reanalysis of these data furnishes several intriguing elements that could raise new questions about the role that DNA methylation plays in the fly genome. The aim of the present paper is to discuss some features that emerge from the analysis of the identified methylated sequences. Received 26 January 2006; received after revision 8 May 2006; accepted 2 June 2006     

转座子的起源及其和物种进化的关系

关于转座子的属性及其起源问题现在还没有解决,在这篇论文中,作者经过分析提出了转座子由病毒或逆转录病毒退化而来的观点,并且阐述了转座子与物种进化之间的关系,主要从转座子影响物种适应性和引起基因突变,染色体畸变两个方面进行了说明。     

The mutator phenotype theory can explain the complex morphology and behaviour of cancers

Almost all solid malignancies exhibit complex cytological and architectural abnormalities, which vary from cell to cell and area to area within the same tumour, and between tumours of the same type. The degrees of these abnormalities do not correlate perfectly with the biological behaviour (especially growth rate and metastatic potential) among the various tumour types. These features of tumours have long been considered to invalidate simple mutational or 'abnormal gene expression' (epigenetic) theories of carcinogenesis. The 'mutator phenotype/clonal selection' hypothesis is based on the now well-established phenomenon of genetic instability of cancer cells, and proposes that this instability is an essential requirement for the development of tumours, and not an irrelevant side-effect of some other process. This paper argues that this hypothesis can provide a satisfactory explanation for the diverse histological and biological features of solid malignancies. Further, because virtually all solid tumours are histologically abnormal, genetic instability is likely to be essential for the malignant process. The concepts of mutator phenotype and clonal selection are therefore supported. Received 8 April 2002; accepted 25 April 2002     

植物转座元件及其表观遗传调控?

植物的转座元件对于植物基因组的组成、进化和基因表达都具有重要影响.植物绝大部分的转座元件都处于沉默状态.植物机体通过一套识别转座元件并进行表观遗传沉默的有效机制,可逆地调控着转座元件的激活与沉默.本文综述了转座元件沉默和激活的表观遗传机制.     

2个棒状杆菌质粒pXZ10142和pXZ10145.1及其转座子TN45的特征分析

对1株棒状杆菌1014中分离到的2个质粒pXZ10145．1和pXZ10142进行了相互关系分析．序列分析显示pXZ10142源自pXZ10145．1．pXZ10145．1包括6个开放读码框(open reading frame,ORF),其中ORF1编码247个氨基酸,经缺失分析是复制酶．在pXZ10145．1上发现1个转座子TN45,转座子上有CMR基因和TNP基因．一对颠倒重复序列IR1a和IR1b位于pXZ10145．1和pXZ10142的交互处,两边是顺向重复序列ATCTAG．     

PiggyBac 转座子在肝脏中的活性研究

本实验运用高压尾静脉注射的方法将PB转座系统导入小鼠肝脏,研究其在肝脏中的转座活性,系统中PB转座子携带表达红色荧光蛋白的基因以指示转座的发生.注射10个月后取出小鼠肝脏,体视荧光显微镜观察肝脏是否有红色荧光,并通过基因组PCR、splinkerette PCR分析PB转座子在肝脏中的插入情况.实验结果表明,PB转座子在肝脏内发生高效转座,检测的68个PB插入位点中有34个位于基因序列,使得PB系统可以作为有效的基因诱变工具来研究基因功能.     

一种酿酒酵母同源重组载体的筛选

将携有mTn-3xHA／LacZ转座子的酿酒酵母文库质粒pHSS6转化大肠杆菌DH5α,挑取单菌落并提取其质粒．将纯化质粒分别用Not Ⅰ酶切后,转化尿嘧啶缺陷型(ura^-)酿酒酵母菌株INVScl,使其同源重组到酿酒酵母基因组中,于SC／ura^-平板上筛选．取长有单菌落酵母的平板进一步筛选重组后上游有强启动子酿酒酵母文库质粒,用近300个质粒转化酵母菌株INVScl,最终得到2株前端有强启动子的酿酒酵母质粒,这2个质粒可作为酿酒酵母同源重组载体广泛应用．     

Asp38与果蝇醇脱氢酶的辅酶特异性

果蝇醇脱氢酶依赖于ＮＡＤ＾＋,Ａｓｐ３８是一个保守残基,它与ＮＡＤ＾＋的ＡＭＰ单位中核糖羟基作用。野生型果蝇ＡＤＨ的Ａｓｐ３８突变成Ａｓｎ３８（Ｄ３８Ｎ）后,Ｋｍ（ａｐｐ）ＮＡＤＰ值下降至１／６２,Ｋｃａｔ／Ｋｍ（ａｐｐ）ＮＡＤＰ值则增加至５９０倍（ＰＨ９．８）。     

水稻转座子Pong特异性RNAi载体TPAS的构建和农杆菌介导的遗传转化条件的优化

为研究水稻转座子Pong的功能,构建了Pong的特异RNAi表达载体TPAS,并利用农杆菌介导法将这个载体转化到水稻的成熟胚愈伤组织,获得了PCR阳性的再生植株.同时通过对农杆菌侵染途径、侵染浓度和侵染时间等的研究,建立并优化了水稻的遗传转化程序,结果表明:在农杆菌侵染成熟胚愈伤组织的时间为3 min,共培养2～4 d...