Large scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression

  title={Large scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression},
  author={Kousaku Okubo and N Hori and Ryo Matoba and T. Niiyama and A Fukushima and Yuko Kojima and Kenichi Matsubara},
  journal={Nature Genetics},
Large scale sequencing of cDNAs provides a complementary approach to structural analysis of the human genome by generating expressed sequence tags (ESTs). We have initiated the large–scale sequencing of a 3′–directed cDNA library from the human liver cell line HepG2, that is a non–biased representation of the mRNA population. 982 random cDNA clones were sequenced yielding more than 270 kilobases. A significant portion of the identified genes encoded secretable proteins and components for… 

Rapid cDNA sequencing (expressed sequence tags) from a directionally cloned human infant brain cDNA library

A human infant brain cDNA library, made specifically for production of expressed sequence tags (ESTs) was evaluated by partial sequencing of over 1,600 clones, and two proteins similar to the Alzheimer's disease amyloid precursor protein were identified.

Fine Expression Profiling of Full-length Transcripts using a Size-unbiased cDNA Library Prepared with the Vector-capping Method

The results suggest that the size-unbiased full-length cDNA library constructed using the vector-capping method will be an ideal resource for fine expression profiling of transcriptional variants with alternative TSSs and alternative splicing.

Identification of 4370 expressed sequence tags from a 3'-end-specific cDNA library of human skeletal muscle by DNA sequencing and filter hybridization.

A new method has been developed for the construction of unbiased cDNA libraries specially designed for the production of ESTs corresponding to the 3'-end portion of the mRNAs, applied to human skeletal muscle, where the analysis of the transcription profile is particularly difficult for the presence of several very abundant transcripts.

Construction of a gene expression profile of a human fetal liver by single-pass cDNA sequencing

An overall gene expression profile of a human fetal liver is obtained by sequencing the 5′ ends of random cDNA clones from an unbiased cDNA library, and many novel genes that might be related to liver growth and hemopoiesis have been identified.

Rapid cDNA sequencing in combination with RNA expression studies in mice identifies a large number of male germ cell-specific sequence tags.

A large number of cDNA clones were isolated from an adult mouse testis cDNA library and partially sequenced. Sequence comparisons revealed that many of them displayed similarities to genes previously

An expression-independent catalog of genes from human chromosome 22.

To accomplish large-scale identification of genes from a single human chromosome, exon amplification was applied to large pools of clones from a flow-sorted human chromosome 22 cosmid library, expected to facilitate fine-structure physical and transcription mapping of human chromosomes, and accelerate the process of disease gene identification.

Semi-multiplex PCR technique for screening of abundant transcripts during systematic sequencing of cDNA libraries.

A very simple method of "semi-multiplex PCR" that allows specific identification of such abundant transcripts before DNA sequencing without using nonrepresentative subtracted libraries is described.



Complementary DNA sequencing: expressed sequence tags and human genome project

Automated partial DNA sequencing was conducted on more than 600 randomly selected human brain complementary DNA (cDNA) clones to generate expressed sequence tags (ESTs), which will facilitate the tagging of most human genes in a few years at a fraction of the cost of complete genomic sequencing.

A novel system for large-scale sequencing of cDNA by PCR amplification.

With this protocol, sample preparation became a non-limiting step, that allowed for large-scale sequencing of cDNA clones and to sequence as many samples as the autosequencer could handle.

Construction of a uniform-abundance (normalized) cDNA library.

Libraries of these fragments are suitable for cDNA subtraction, screening, or selection by hybridization and make it possible to detect and analyze cDNA corresponding to species of mRNA present at a low level in a small fraction of the cells in a complex tissue.

Sequence identification of 2,375 human brain genes

2,672 new, independent cDNA clones isolated from four human brain cDNA libraries are partially sequenced to generate 2,375 expressed sequence tags to nuclear-encoded genes, representing an approximate doubling of the number of human genes identified by DNA sequencing and may represent as many as 5% of the genes in the human genome.

Characterization of cDNA clones for the human c-yes gene

Three c-yes cDNA clones obtained from poly(A)+ RNA of human embryo fibroblasts showed that they contained inserts corresponding to nearly full-length human c- yes mRNA, which could encode a polypeptide of 543 amino acids with a relative molecular weight (Mr) of 60,801.

High-efficiency cloning of full-length cDNA

The cloning procedure described here mitigates this shortcoming by attributing the high efficiency of cloning full- or nearly full-length cDNA to the fact that the plasmid DNA vector itself serves as the primer for first- and second-strand cDNA synthesis.

An 'equalized cDNA library' by the reassociation of short double-stranded cDNAs.

  • M. S. Ko
  • Biology, Chemistry
    Nucleic acids research
  • 1990
A library of mouse fibroblastoid Ltk- cells with nearly equal representations of cDNA clones is tried to make, indicating the usefulness of the current procedure for making equalized cDNA libraries.

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

The testis enzyme contains the second of the two putative metal-binding sites (His-Glu-Met-Gly-His) identified in endothelial ACE, which indicates that the functionally active catalytic site is within the C-terminal domain of the endothelial enzyme, accounting for the previous finding that these two structurally dissimilar isozymes are virtually identical catalytically.