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。     

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.     

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     

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．     

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...     

利用转座子Tn917构建杀虫Bt工程菌

利用链球菌（Ｓｔｒｅｐｔｏｃｏｃｕｓｆａｅｃａｌｉｓ）的转座子Ｔｎ９１７衍生载体ｐＴＶ１Ｔｓ将苏云金杆菌（Ｂａｃｉｌｕｓｔｈｕｒｉｎｇｉｅｎｓｉｓ,Ｂｔ）的２个杀虫晶体蛋白（ＩＣＰｓ）基因整合进Ｂｔ染色体上,转座频率达５９×１０－５．将对双翅目昆虫有毒效的ＩＣＰｓ基因ｃｒｙ１１Ａ和广谱溶细胞且对蚊子等有毒效的ｃｙｔ１Ａ基因分别克隆到ｐＴＶ１Ｔｓ上,用高压电激法转入对鳞翅目昆虫有毒效的库斯塔克亚种（Ｂｔｓｕｂｓｐｋｕｒｓｔａｋｉ）ＨＤ１菌株,分别筛选到在染色体基因组上整合进这２个基因的突变株ＨＴ２６和ＨＴ４５,整合基因均能在这２株工程菌中获得表达并形成正常的伴孢晶体．生物测定结果显示,表达的Ｃｒｙ１１Ａ和Ｃｙｔ１Ａ晶体蛋白有很高的杀虫活性,对库蚊三龄幼虫的ＬＣ５０分别为８３ｎｇ／ｍＬ和６４ｎｇ／ｍＬ,同时Ｃｙｔ１Ａ晶体蛋白对培养的昆虫细胞有较强的溶解作用．其结果为创造新型基因工程苏云金杆菌杀虫剂提供了新的重要途径     

Construction and genetic analysis of mutator insertion mutant population in maize

A total of 26718 M1 plants were ob- tained by crossing the active mutator transposon donor parents (Q105, WW51, 115F, V26-2 and 919J) with the recipient parents (Hz85,W328 with Bz gene and S-Mo17Rf3Rf3). The phenotypes of M1 plants were observed in the field. M1 plants were self-pollinated to develop the mutator insertion-mutagenized M2 seeds. The transposition frequency of the mutator in the genome was calculated based on the spotted aleurone phenotype of the M2 seeds. The results showed that: (1) the mutation frequency of M1 phe- notypes in the field was 0.07 in the population of W328×Mu; (2) the mutation frequency of spotted aleurone seeds on the M2 ears was 0.122 in the population of W328×Mu; (3) five S-cytoplasm male-sterile plants were found among 22500 M1 plants of S-Mo17Rf3Rf3×Mu, with the transposition frequency about 2.2×10?4 per locus. 99 flanking se- quences of mutator transposition were amplified by the modified MuTAIL-PCR, and 59 non-redundant sequences with length around 400 bp were obtained. After bioinformatic analysis, 27 sequences of them could be annotated, using non-redundant nucleotide database of maize, rice, and Arabidopsis. 36 se- quences of them were located on the genetic map of maize by comparative genomics, and several flank- ing sequences of mutator insertion were mapped on the single marker locus. Hotspot sequences of mu- tator transposition were revealed by comparing the homologies between the 9-bp target site duplication of the mutator insertion. The putative functions of 8 flanking sequences of mutator transposition had identity with the functions of their correspondingmarker. The constructed mutator insertion mutant population in maize will facilitate the new gene discovery and functional genomics study in maize.