ORGANIZATION AND FUNCTION OF A HUMAN REPETITIVE SEQUENCE

一个低重复的2.2 kb序列已从人类基因组中分离出来。 根据初步研究，我们怀疑这个序列是一个移动的或 转座因子 本提案的主要目的是确认这一点 假设，并发现这个序列是否是一个部分， 调节其他基因的表达。 为此，我们提议： 1)对克隆的2.2kb重复DNA元件和相关成员进行测序 它的家人。 M13克隆和桑格双脱氧”测序程序将 用于获得将被分析用于开放阅读的核苷酸数据 框架、保守序列和其他类似于逆转录病毒的特征 和转座因子。 2)来研究这种重复的组织 家族，因为它出现在人类基因组的各个部分。 Lambda 将筛选人12号染色体文库中的重组体， 与2.2kb序列杂交。 来自这些重组体的DNA将被 与2.2kb序列相比，在组织和程度方面， 交叉杂交 3)研究重复家庭的转录本， 来自各种细胞和组织的RNA的北方印迹分析， 筛选预先存在的cDNA文库。 RNA的北方印迹分析 不同的来源将为我们提供有关大小和类型的信息， 成绩单 分析含有同源序列的cDNA克隆， 2.2kb片段将揭示是否单独的重复元件 转录或仅与其他基因结合。 4)探索使用 2.2kb重复序列家族的一部分作为遗传研究的标记。 家庭成员将用于人类12号染色体的区域定位 和21，并进一步扩展到小缺失的检测， 限制性片段长度多态性。 这些研究将导致 关于一类重复序列的关系的信息 人类基因组的调节、分化和发育。 还有， 这些克隆的重复元件可能在医学上有应用， 作为遗传标记用于检测遗传和发育 疾病，包括癌症。

A low-repetitive 2.2 kb sequence has been isolated from the human genome. Based on preliminary studies, we suspect that this sequence is a mobile or transposable element. The major aim of this proposal is to confirm this hypothnesis and to discover whether or not this sequence is a part of or regulates the expression of other genes. Toward this end, we propose: 1) to sequence a cloned 2.2 kb repetitive DNA element and related members of its family. M13 cloning and Sanger's dideoxy" sequencing procedure will be used to obtain nucleotide data which will be analyzed for open-reading frames, conserved sequence, and other features which resemble retroviruses and transposable elements. 2) to study the organization of this repeat family as it appears in various portions of the human genome. Lambda libraries of human chromosome 12 will be screened for recombinants which hybridize to the 2.2 kb sequence. DNA from such recombinants will be compared to the 2.2 kb sequence in terms of organization and degree of cross-hybridization. 3) to study transcripts of the repeat family by Northern blot analysis of RNA from various cells and tissues and by screening preexisting cDNA libraries. Northern blot analysis of RNA from different sources will give us information on the sizes and types of transcripts. Analysis of cDNA clones containing sequences homologous to the 2.2 kb fragment will reveal whether the repetitive element alone is transcribed or only in conjunction with other genes. 4) to explore the use of the 2.2 kb repetitive sequence family as markers for genetic studies. Family members will be used for regional mapping of human chromosomes 12 and 21 and further extended to the detection of small deletions and restriction fragment length polymorphisms. These studies will result in information regarding the relationship of a class of repetitive sequences to regulation, differentiation and development in the human genome. Also, these cloned repetitive elements mayt have application to medicine in their use as genetic markers for the detection of genetic and developmental diseases, including cancer.