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The reduced expression of 6Ckine in the plt mouse results from the deletion of one of two 6Ckine genes.

Vassileva G, Soto H, Zlotnik A, Nakano H, Kakiuchi T, Hedrick JA, Lira SA - J. Exp. Med. (1999)

Bottom Line: 6Ckine is an unusual chemokine capable of attracting naive T lymphocytes in vitro.It has been recently reported that lack of 6Ckine expression in lymphoid organs is a prominent characteristic of mice homozygous for the paucity of lymph node T cell (plt) mutation.These mice show reduced numbers of T cells in lymph nodes, Peyer's patches, and the white pulp of the spleen.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.

ABSTRACT
6Ckine is an unusual chemokine capable of attracting naive T lymphocytes in vitro. It has been recently reported that lack of 6Ckine expression in lymphoid organs is a prominent characteristic of mice homozygous for the paucity of lymph node T cell (plt) mutation. These mice show reduced numbers of T cells in lymph nodes, Peyer's patches, and the white pulp of the spleen. The genetic reason for the lack of 6Ckine expression in the plt mouse, however, has remained unknown. Here we demonstrate that mouse 6Ckine is encoded by two genes, one of which is expressed in lymphoid organs and is deleted in plt mice. A second 6Ckine gene is intact and expressed in the plt mouse.

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(A) The 6Ckine genes found on the two BACs are distinct. A schematic of the sequence of SacI fragments (7.5 kb) from 6CKBAC1 (leu) and 6CKBAC2 (ser) is shown. Black and white boxes represent 6Ckine exons. Open triangles indicate larger deletions. Single base differences outside of the exons are not shown. The four nucleotide differences identified in coding regions (black boxes) and noncoding regions (white box) of the two 6Ckine genes are shown. Vertical arrows identify the amino acid difference at position 65. Horizontal arrows indicate the positions of the PCR primers used for analysis of 6Ckine gene locus. nt, nucleotide. (B) Sequences of the intron–exon boundaries are identical between the two copies with the exception of the intron2–exon3 junction. The 6Ckine-ser sequence is added to the intron2–exon3 boundary sequence to illustrate the single nucleotide difference (arrow). These sequence data are available from EMBL/GenBank/DDBJ under accession numbers AF171085 and AF171086 for 6Ckine-leu and 6Ckine-ser, respectively.
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Figure 2: (A) The 6Ckine genes found on the two BACs are distinct. A schematic of the sequence of SacI fragments (7.5 kb) from 6CKBAC1 (leu) and 6CKBAC2 (ser) is shown. Black and white boxes represent 6Ckine exons. Open triangles indicate larger deletions. Single base differences outside of the exons are not shown. The four nucleotide differences identified in coding regions (black boxes) and noncoding regions (white box) of the two 6Ckine genes are shown. Vertical arrows identify the amino acid difference at position 65. Horizontal arrows indicate the positions of the PCR primers used for analysis of 6Ckine gene locus. nt, nucleotide. (B) Sequences of the intron–exon boundaries are identical between the two copies with the exception of the intron2–exon3 junction. The 6Ckine-ser sequence is added to the intron2–exon3 boundary sequence to illustrate the single nucleotide difference (arrow). These sequence data are available from EMBL/GenBank/DDBJ under accession numbers AF171085 and AF171086 for 6Ckine-leu and 6Ckine-ser, respectively.

Mentions: A pair of PCR primers GV104 (5′-GTA GAC CTA GAG AGT CAG AAG-3′) and GV125 (5′-CGC GGA TCC TTG GAG GAG GAA CCA CAG T-3′), shown in Fig. 2, were used to amplify 1.35- and 1.2-kb fragments that included part of the 3′ untranslated region (UTR) and ∼1 kb downstream of the gene. PCR conditions were: 94°C for 2 min; 25 cycles of 94°C for 30 s, 60°C for 30 s, 72°C for 1 min; and 72°C for 5 min. Both fragments were subcloned into pCR2.1 TA cloning vector (Invitrogen Corp.) and sequenced. Another pair of PCR primers, GV95 (5′-ATG GCT CAG ATG ATG ACT CT-3′) and GV105 (5′-CTT CTG ACT CTC TAG GTC TAC-3′) (see Fig. 2) were used to amplify the 5′ coding region and part of the 3′ UTR of the 6Ckine gene (899 bp) from wild-type and BALB/c-plt (10th backcrossed generation) tail DNA. These fragments were also subcloned into pCR2.1 TA cloning vector (Invitrogen Corp.) and sequenced.


The reduced expression of 6Ckine in the plt mouse results from the deletion of one of two 6Ckine genes.

Vassileva G, Soto H, Zlotnik A, Nakano H, Kakiuchi T, Hedrick JA, Lira SA - J. Exp. Med. (1999)

(A) The 6Ckine genes found on the two BACs are distinct. A schematic of the sequence of SacI fragments (7.5 kb) from 6CKBAC1 (leu) and 6CKBAC2 (ser) is shown. Black and white boxes represent 6Ckine exons. Open triangles indicate larger deletions. Single base differences outside of the exons are not shown. The four nucleotide differences identified in coding regions (black boxes) and noncoding regions (white box) of the two 6Ckine genes are shown. Vertical arrows identify the amino acid difference at position 65. Horizontal arrows indicate the positions of the PCR primers used for analysis of 6Ckine gene locus. nt, nucleotide. (B) Sequences of the intron–exon boundaries are identical between the two copies with the exception of the intron2–exon3 junction. The 6Ckine-ser sequence is added to the intron2–exon3 boundary sequence to illustrate the single nucleotide difference (arrow). These sequence data are available from EMBL/GenBank/DDBJ under accession numbers AF171085 and AF171086 for 6Ckine-leu and 6Ckine-ser, respectively.
© Copyright Policy
Related In: Results  -  Collection

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Figure 2: (A) The 6Ckine genes found on the two BACs are distinct. A schematic of the sequence of SacI fragments (7.5 kb) from 6CKBAC1 (leu) and 6CKBAC2 (ser) is shown. Black and white boxes represent 6Ckine exons. Open triangles indicate larger deletions. Single base differences outside of the exons are not shown. The four nucleotide differences identified in coding regions (black boxes) and noncoding regions (white box) of the two 6Ckine genes are shown. Vertical arrows identify the amino acid difference at position 65. Horizontal arrows indicate the positions of the PCR primers used for analysis of 6Ckine gene locus. nt, nucleotide. (B) Sequences of the intron–exon boundaries are identical between the two copies with the exception of the intron2–exon3 junction. The 6Ckine-ser sequence is added to the intron2–exon3 boundary sequence to illustrate the single nucleotide difference (arrow). These sequence data are available from EMBL/GenBank/DDBJ under accession numbers AF171085 and AF171086 for 6Ckine-leu and 6Ckine-ser, respectively.
Mentions: A pair of PCR primers GV104 (5′-GTA GAC CTA GAG AGT CAG AAG-3′) and GV125 (5′-CGC GGA TCC TTG GAG GAG GAA CCA CAG T-3′), shown in Fig. 2, were used to amplify 1.35- and 1.2-kb fragments that included part of the 3′ untranslated region (UTR) and ∼1 kb downstream of the gene. PCR conditions were: 94°C for 2 min; 25 cycles of 94°C for 30 s, 60°C for 30 s, 72°C for 1 min; and 72°C for 5 min. Both fragments were subcloned into pCR2.1 TA cloning vector (Invitrogen Corp.) and sequenced. Another pair of PCR primers, GV95 (5′-ATG GCT CAG ATG ATG ACT CT-3′) and GV105 (5′-CTT CTG ACT CTC TAG GTC TAC-3′) (see Fig. 2) were used to amplify the 5′ coding region and part of the 3′ UTR of the 6Ckine gene (899 bp) from wild-type and BALB/c-plt (10th backcrossed generation) tail DNA. These fragments were also subcloned into pCR2.1 TA cloning vector (Invitrogen Corp.) and sequenced.

Bottom Line: 6Ckine is an unusual chemokine capable of attracting naive T lymphocytes in vitro.It has been recently reported that lack of 6Ckine expression in lymphoid organs is a prominent characteristic of mice homozygous for the paucity of lymph node T cell (plt) mutation.These mice show reduced numbers of T cells in lymph nodes, Peyer's patches, and the white pulp of the spleen.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.

ABSTRACT
6Ckine is an unusual chemokine capable of attracting naive T lymphocytes in vitro. It has been recently reported that lack of 6Ckine expression in lymphoid organs is a prominent characteristic of mice homozygous for the paucity of lymph node T cell (plt) mutation. These mice show reduced numbers of T cells in lymph nodes, Peyer's patches, and the white pulp of the spleen. The genetic reason for the lack of 6Ckine expression in the plt mouse, however, has remained unknown. Here we demonstrate that mouse 6Ckine is encoded by two genes, one of which is expressed in lymphoid organs and is deleted in plt mice. A second 6Ckine gene is intact and expressed in the plt mouse.

Show MeSH
Related in: MedlinePlus