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Title: Nucleotide sequences of three distinct clones coding for rat heavy chain class 1 major hitocompatibility antigens

Abstract

Poly(A){sup +} RNAs were isolated from ConconavalinA stimulated splenocytes of BUF (RT1.A{sup b}), PVG (RT1.A{sup c}), or PVG.1U (RT1.A{sup u}) rats, respectively, using a Micro-Fast Track kit. After reverse transcription with a synthetic oligo-d(T) primer (5{sup {prime}}-CAT GAT CGA ATT CAC GCG TCT AGA TTT TTT TTT TTT TTT TTT TTT TTT TVN-3{sup {prime}}, V = A+G+C, N = A+T+G+C; Genosys, Woodland, TX), 1.6 kilobase products, which encode the entire MHC class I protein and the 3{sup {prime}} non-translated region including the poly-A tail, were amplified by polymerase chain reaction (PCR) using two synthetic oligonucleotide primers (Genosys). The upstream primer (5{sup {prime}}-GTC CGG GWT CTC AGA TGG GG C-3{sup {prime}}, W = A+T) was designed based upon the published rat class I sequences of eight genes: RT1.1{sup a} M31018; rat LW2 gene X70066; RT1.1{sup 1}, L26224 X79719; RT1.A{sup u} X82669, and RT1.Aw3 L40363, RT1.E{sup u} L40365, RT1.C{sup 1} L40362. The downstream primer (5{sup {prime}}) ATG ATC GAA TTC ACG CGT CTA GA-3{sup {prime}} was the portion of the oligo-d(T) primer used for reverse transcription. The purified PCR products were inserted into pCR II cloning vectors (Invitrogen). Automated sequencing of plasmid cDNAs from the positive clones obtained from three repeated PCRmore » amplifications identified by restriction enzyme mapping were reproducible. Comparison between new sequences of the heavy chain class I genes and those available in GenBank. 7 refs., 1 fig.« less

Authors:
; ;  [1]
  1. Univ. of Texas Medical School, Houston, TX (United States); and others
Publication Date:
OSTI Identifier:
273489
Resource Type:
Journal Article
Journal Name:
Immunogenetics
Additional Journal Information:
Journal Volume: 43; Journal Issue: 5; Other Information: PBD: 1996
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; DNA SEQUENCING; RELIABILITY; ANTIGENS; GENES; HISTOCOMPATIBILITY COMPLEX; DNA-CLONING; NUCLEOTIDES; RATS

Citation Formats

Wang, M, Stepkowski, S M, and Tain, L. Nucleotide sequences of three distinct clones coding for rat heavy chain class 1 major hitocompatibility antigens. United States: N. p., 1996. Web. doi:10.1007/s002510050069.
Wang, M, Stepkowski, S M, & Tain, L. Nucleotide sequences of three distinct clones coding for rat heavy chain class 1 major hitocompatibility antigens. United States. https://doi.org/10.1007/s002510050069
Wang, M, Stepkowski, S M, and Tain, L. 1996. "Nucleotide sequences of three distinct clones coding for rat heavy chain class 1 major hitocompatibility antigens". United States. https://doi.org/10.1007/s002510050069.
@article{osti_273489,
title = {Nucleotide sequences of three distinct clones coding for rat heavy chain class 1 major hitocompatibility antigens},
author = {Wang, M and Stepkowski, S M and Tain, L},
abstractNote = {Poly(A){sup +} RNAs were isolated from ConconavalinA stimulated splenocytes of BUF (RT1.A{sup b}), PVG (RT1.A{sup c}), or PVG.1U (RT1.A{sup u}) rats, respectively, using a Micro-Fast Track kit. After reverse transcription with a synthetic oligo-d(T) primer (5{sup {prime}}-CAT GAT CGA ATT CAC GCG TCT AGA TTT TTT TTT TTT TTT TTT TTT TTT TVN-3{sup {prime}}, V = A+G+C, N = A+T+G+C; Genosys, Woodland, TX), 1.6 kilobase products, which encode the entire MHC class I protein and the 3{sup {prime}} non-translated region including the poly-A tail, were amplified by polymerase chain reaction (PCR) using two synthetic oligonucleotide primers (Genosys). The upstream primer (5{sup {prime}}-GTC CGG GWT CTC AGA TGG GG C-3{sup {prime}}, W = A+T) was designed based upon the published rat class I sequences of eight genes: RT1.1{sup a} M31018; rat LW2 gene X70066; RT1.1{sup 1}, L26224 X79719; RT1.A{sup u} X82669, and RT1.Aw3 L40363, RT1.E{sup u} L40365, RT1.C{sup 1} L40362. The downstream primer (5{sup {prime}}) ATG ATC GAA TTC ACG CGT CTA GA-3{sup {prime}} was the portion of the oligo-d(T) primer used for reverse transcription. The purified PCR products were inserted into pCR II cloning vectors (Invitrogen). Automated sequencing of plasmid cDNAs from the positive clones obtained from three repeated PCR amplifications identified by restriction enzyme mapping were reproducible. Comparison between new sequences of the heavy chain class I genes and those available in GenBank. 7 refs., 1 fig.},
doi = {10.1007/s002510050069},
url = {https://www.osti.gov/biblio/273489}, journal = {Immunogenetics},
number = 5,
volume = 43,
place = {United States},
year = {Sun Sep 01 00:00:00 EDT 1996},
month = {Sun Sep 01 00:00:00 EDT 1996}
}