NIA Mouse Osteoblast cDNA Library (Long)

Library Description

Organism:	Mus musculus
Strain:	C3H/He
Tissue:	Osteoblast
Cell line:	KUSA-A1 cells
Vector:	pSPORT1 (Invitrogen)
Vector type:	plasmid (ampicillin resistant)
Cloning site 1:	SalI
Cloning site 2:	NotI
Lab host:	DH10B

Mouse cDNA project by the Laboratory of Genetics, National Institute on Aging (NIA), Intramural Research Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a long-transcript enriched cDNA library (Ref. Genome Res. 11: 1553-1558 (2001). [PMID: 11544199]). Total RNAs were obtained from Dr. Akihiro Umezawa (Keio University School of Medicine, Japan). Double-stranded cDNAs were synthesized with an Oligo(dT) primer [Invitrogen: 5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 2.1 ug of total RNA, treated with T4 DNA polymerase, and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal4, purified by phenol/chloroform, and separated from free linkers by Centricon 100. Then, the cDNAs were amplified by long-range high fidelity PCR using Ex Taq polymerase (Takara) with a primer Sal4-S. The products were purified by phenol/chloroform and Centricon 100. The cDNAs were digested with SalI and NotI enzymes and cloned into SalI/NotI site of pSPORT1 plasmid vector. The DH10B E. coli host was transformed with the ligation mixture by the standard chemical method. The average insert size is about 3.0 kb. The library was constructed by Yulan Piao (NIA).

Source: Dawood B. Dudekula, National Institute on Aging/National Institutes of Health

Gene Content Analysis

7,696 ESTs from this library were grouped into 1,960 UniGene entries (putative genes) [UniGene build #194, 25-Jan-2013]. EST counts for each entry may be used to calculate an approximate expression level in transcripts per million (TPM).

ESTsTPM UniGene Entry

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268	34823	Mm.4159	Thrombospondin 1.
227	29496	Mm.277792	Collagen, type I, alpha 2.
195	25338	Mm.277735	Collagen, type I, alpha 1.
80	10395	Mm.287146	Fibromodulin.
75	9745	Mm.138471	Eukaryotic translation elongation factor 1 alpha 1.
60	7796	Mm.326799	Eukaryotic translation elongation factor 2.
55	7147	Mm.268000	Vimentin.
54	7017	Mm.2608	Biglycan.
47	6107	Mm.291442	Secreted acidic cysteine rich glycoprotein.
46	5977	Mm.391967	Actin, beta.
46	5977	Mm.16660	Prolyl 4-hydroxylase, beta polypeptide.
39	5068	Mm.419997	Ubiquitin C.
38	4938	Mm.10299	Collagen, type V, alpha 2.
37	4808	Mm.193099	Fibronectin 1.
32	4158	Mm.249555	Collagen, type III, alpha 1.
31	4028	Mm.265610	Polypyrimidine tract binding protein 1.
29	3768	Mm.336743	Heat shock protein 8.
29	3768	Mm.243014	Lamin A.
28	3638	Mm.392113	Tubulin, alpha 1B.
28	3638	Mm.379939	Transcribed locus, strongly similar to NP_989488.2 EEF1A1 gene product [Gallus gallus].
28	3638	Mm.290774	Heat shock protein 8.
27	3508	Mm.2180	Heat shock protein 90 alpha (cytosolic), class B member 1.
26	3378	Mm.7281	Collagen, type V, alpha 1.
26	3378	Mm.4871	Tissue inhibitor of metalloproteinase 3.
26	3378	Mm.21109	Gelsolin.
25	3248	Mm.87773	Heat shock protein 90, beta (Grp94), member 1.
25	3248	Mm.330160	Heat shock protein 5.
25	3248	Mm.24174	Alanyl-tRNA synthetase.
25	3248	Mm.22130	Ribophorin II.
21	2729	Mm.289915	Damage specific DNA binding protein 1.
20	2599	Mm.295533	Filamin, alpha.
20	2599	Mm.289936	Phosphoserine aminotransferase 1.
19	2469	Mm.426706	Actin, gamma, cytoplasmic 1.
19	2469	Mm.326167	Pyruvate kinase, muscle.
19	2469	Mm.30012	High density lipoprotein (HDL) binding protein.
18	2339	Mm.335315	Eukaryotic translation elongation factor 1 alpha 1.
17	2209	Mm.4114	Solute carrier family 3 (activators of dibasic and neutral amino acid transport), member 2.
17	2209	Mm.3894	FK506 binding protein 10.
17	2209	Mm.296985	Chaperonin containing Tcp1, subunit 4 (delta).
17	2209	Mm.29324	Lactate dehydrogenase A.
17	2209	Mm.273538	Tubulin, beta 5 class I.
16	2079	Mm.491096	DEAD (Asp-Glu-Ala-Asp) box polypeptide 5.
16	2079	Mm.427589	Cell cycle associated protein 1.
16	2079	Mm.392075	Calumenin.
16	2079	Mm.2942	Asparagine synthetase.
16	2079	Mm.28375	Sec61 alpha 1 subunit (S. cerevisiae).
16	2079	Mm.277498	Prosaposin.
16	2079	Mm.271967	LIM and SH3 protein 1.
16	2079	Mm.245976	Valosin containing protein.
16	2079	Mm.22776	Eukaryotic translation initiation factor 3, subunit C.
16	2079	Mm.168478	Cytoskeleton associated protein 5.
15	1949	Mm.88212	Tubulin, alpha 1C.
15	1949	Mm.454219	Tenascin C.
15	1949	Mm.3196	Alpha glucosidase 2 alpha neutral subunit.
15	1949	Mm.279923	Neural precursor cell expressed, developmentally down-regulated 4.
15	1949	Mm.234502	Myosin IC.
14	1819	Mm.485669	Eukaryotic translation elongation factor 1 alpha 1.
14	1819	Mm.196173	Transcribed locus, strongly similar to NP_033739.1 actin, cytoplasmic 2 [Mus musculus].
13	1689	Mm.426664	Corneodesmosin.
13	1689	Mm.30158	Fatty acid desaturase 1.
13	1689	Mm.28270	Cell division cycle 5-like (S. pombe).
13	1689	Mm.252080	Phosphogluconate dehydrogenase.
13	1689	Mm.2509	Collagen, type VI, alpha 1.
13	1689	Mm.234912	Plectin.
13	1689	Mm.222867	Death-associated protein.
13	1689	Mm.221452	Importin 5.
13	1689

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Monday, June 30, 2014

KUSA-A1 データベース NCBI NIA Mouse Osteoblast cDNA Library (Long)

NIA Mouse Osteoblast cDNA Library (Long)

Library Description

Gene Content Analysis

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