Cloning And Characterization Of Two Novel Genes Encoding Zinc Finger Proteins: HZF1 And HZF2

Zinc finger domain is one of the important DNA binding motifs. A lot of transcriptional factors have zinc finger DNA binding domains. According to the conserved amino acids in zinc finger domain we can design degenerated primers to amplify zinc finger ESTs with the cDNA template from cell lines or tissues. Then we can isolate and clone zinc finger protein encoding gene by screening cDNA library. Human erythroleukemia cell line(HEL) can be induced to express globin by hemin and undergoes-a drastic macrophage-like shift with PMA. There must be some transcriptional factors with zinc finger motifs involved in the process of the differentiations.We first induced HEL cell line by hemin and PMA respectively. Then amplified cDNA with the zinc finger degenerated primers. In our study, we acquired 22 novel zinc finger expressed seqence tags(ESTs), 5 of them are ESTs of known genes encoding zinc finger proteins, 17 of them are new ESTs (GenBank accession number: BE240813~BE240828). Two EST(One come from hemin induced HEL cells while the other come from PMA induced HEL cells) were chosen as the probes for cDNA library screening and several full length cDNA clones were isolated from human bone marrow cDNA library. In this thesis, the cloning and characterization of two full-length cDNAs were reported.HZF1 gene (EST was acquired from hemin induced HEL cells, GenBank accession number: AF244088) is a full-length cDNA isolated from human bone marrow cDNA library. The cDNA contains a 2061 bp open reading frame encoding a protein of 686 amino acids. In the 46-48nt there is a ATG start codon. The deduced protein contains 671 amino acids. There are 15 typical C2H2 zinc finger motifs and 2 C2RH zinc finger motifs. HZF1 carrying several potential modification sites such as N-myristoylation, casein kinase II- and protein kinase c-phosphorylation. HZF1 has one transcript, ~2.5 kb, shown by multiple tissue...