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- Title
Molecular cloning and expression of two chicken invariant chain isoforms produced by alternative splicing.
- Authors
Zhong, Dalian; Yu, Weiyi; Liu, Yuhua; Liu, Jing; Li, Jinnian
- Abstract
The biosynthesis of distinct forms of the invariant chain (Ii) protein from a unique gene as the result of differential splicing patterns has been observed in humans and mice. However, there have been no reports on the existence of Ii isoforms in avian species. In the present study, we identified two chickenIicDNAs by RT-PCR and RACE, and examined theIigene copy number, mRNA expression and protein expression by Southern blotting, Northern blotting and immunofluorescence confocal microscopy, respectively. One of theIicDNAs, namedIi-1, was 1,151 bp in length, and had an open reading frame (ORF) of 672 nucleotides, in agreement with a previously identified chickenIisequence; the other, namedIi-2, was 1,337 bp long and had an ORF of 861 nucleotides. Southern blotting confirmed that these cDNAs were derived from a single copy gene. Northern blotting performed with total RNA from various tissues of 6-week-old chickens revealed high levels ofIi-1andIi-2mRNA expression in the spleen and bursa of Fabricius, and low levels ofIi-1expression in the thymus, heart and liver, whileIi-2was not expressed in these tissues. High levels of expression of bothIiisoforms were detected in the spleen and bursa of Fabricius during late embryogenesis. Immunofluorescence staining showed that Ii proteins were expressed in the cell membranes of the splenocytes. These data suggest that chicken Ii exists in two isoforms resulting from alternative splicing, and is strongly expressed in the major immune organs.
- Subjects
MOLECULAR cloning; MOLECULAR genetics; BIOSYNTHESIS; BIOCHEMICAL engineering; IMMUNOGLOBULINS; IMMUNOFLUORESCENCE
- Publication
Immunogenetics, 2004, Vol 56, Issue 9, p650
- ISSN
0093-7711
- Publication type
Article
- DOI
10.1007/s00251-004-0726-6