CDNAs libraries

Fig. 4. Steps in making a cDNA library. Cellular mRNA is used as a template to make a complementary DNA. This cDNA is then ligated to a plasmid,...

Homology screening. Using oligonucleotide probes based on known receptor sequences, search cDNA libraries for homologous sequences which may code for related receptors. The clones are then isolated and sequenced and used in expression studies to confirm the identity of the receptor. 

The PE2 isoform has been purified and fully sequenced (Markovic and Joumval 1986). Using this sequence data Ray et al (1988) succeeded in isolating a clone fi om a tomato fruit cDNA library. The predicted amino acid sequence fi om this clone had high homology to the actual amino acid sequence determined for PE2 but was not identical. Subsequent screening of the tomato... 

A.aculeatus CBS 115.80 was grown on apple pectin in a fermenter and strong induction of Rgase production could be observed. Total RNA was isolated from such an RGase producing culture and used for the construction of a cDNA library. 

The first Ca -ATPase clones were isolated by probing cDNA libraries with radiolabeled synthetic oligonucleotides [42] that represented an established amino acid sequence ((Trp-) Phe Met Tyr Ala) in the fast-twitch skeletal muscle... 

The amino acid sequence of the human erythrocyte glucose transporter was deduced from the nucleotide sequence of a cDNA clone in 1985 [106]. Polyclonal antibodies raised against the protein were used to screen a Xgtl I cDNA library prepared from the human hepatocellular carcinoma cell line HepG2. (Like many other transformed... 

A very similar cloning strategy has been used for the 48-kDa subunit of the glycine-receptor channel by Betz and collaborators [4]. Strychnine was used as an affinity ligand, and synthetic oligopeptides were constructed from the purified material. They have been used to screen cDNA libraries. 

Isoforms of the oi, subunit have now also been cloned from cardiac [80] and smooth muscle [81,82] cDNA libraries, and partial clones resembling this structure have been obtained from brain libraries [20]. In addition, a cDNA encoding a high... 

The murine stromal-derived factor-la (SDF-1)/CXCL12 was originally cloned from a cDNA library derived from the bone marrow stromal cell tine ST2 by a method to clone cDNAs that carry specific amino terminal signal sequences, such as those encoding intercellular signaling-transducing molecules... 

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