Structure and expression of the Lyt-3 agene of C.AKR mice

The mouse Lyt-3 ^agene, which encodes the Lyt-3.1 T-cell surface alloantigen of the C.AKR strain, has been cloned, and the nucleotide sequence of its exons and more than 2 kb of 5 flanking sequence have been determined. The gene extends over approximately 16 kb of DNA and consists of six exons encoding leader, leader plus V-like domain, membrane-proximal, transmembrane, and cytoplasmic domains. The only difference between the coding region of the Lyt-3 ^agene and the cDNA sequences reported for Lyt-3 ^b(Nakauchi et al. 1987, Panaccio et al. 1987) is at position 77 of the mature protein where Lyt-3 ^aencodes serine and Lyt-3 ^bencodes arginine. This substitution must therefore be the basis for the serological distinction between the Lyt-3.1 and Lyt-3.2 alloantigens. Potential TATA and CAAT sequences, two Sp1 protein binding sites, two extended repeats of the dinucleotide, CA, a number of short inverted repeats, and an inverted segment of the mouse B1 repetitive sequence are found 5 to the Lyt-3 ^agene. Two consensus poly-A addition signals and a complete copy of the mouse B1 sequence are found 3 to the gene. Both B1-related regions are flanked by short direct repeats suggesting that they arose by an insertional mechanism. Cotransfection of the Lyt-3 ^agene together with a cloned Lyt-2 ^agene resulted in expression of both Lyt-2 and Lyt-3.1 on the surface of Ltk^– and BW5147 cells. Transfection of the Lyt-3 ^agene without Lyt-2 ^aled to expression of Lyt-3-related cellular RNA but did not result in surface expression of Lyt-3.1, suggesting that the Lyt-3 glycoprotein is not expressed on the cell surface in the absence of Lyt-2.     

Construction and properties of new Lyt-congenic strains and anti-Lyt-2.2 and anti-Lyt-3.1 monoclonal antibodies

Hybridomas producing mouse monoclonal IgM antibodies specific for Lyt-2.2 and Lyt-3.1 T-cell surface alloantigens have been constructed. Cytotoxic titers of ascites fluids were found to be 10^–6 or greater and no lysis of thymocytes of congenic strains bearing the alternative allele was observed at the lowest dilutions tested (12). The anti-Lyt-2.2 monoclonal antibody (HO-2.2) specifically precipiated from extracts of Lyt-2.2-positive thymocytes molecular species indistinguishable from those precipitated by conventional anti-Lyt-2.2 sera. However, by immunoprecipitation criteria (though not by cytotoxicity), the anti-Lyt-3.1 antibody (HO-3.1) demonstrated some cross-reactivity with similar molecular species from Lyt-3.1-negative thymocytes.In addition, three new strains of mice differing from existing strains in the region of theLyt-2 and4Lyt-3 loci have been constructed. They are: C.C58-Lyt-2^a, Lyt-3^a and C.AKR-Lyt-2^a, Lyt-3^a, congenic with Balb/cAn and bearingLyt-2 ^a andLyt-3 ^a alleles of C58/J and AKR/J, respectively; and AKR.C-Lyt-2^b, Lyt-3^b, congenic with AKR/J and bearing theLyt-2 ^b andLyt-3 ^b alleles of Balb/cJ.Abbreviations used in this paper DMSO dimethylsulfoxide - NP40 Nonidet P-40 detergent - SaCI Staphylococcus aureus, Cowan I strain - SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoreses - NP-NET buffer 0.15 M NaCl, 0.005 M EDTA, 0.05 M Tris, 0.02% sodium azide, pH 7.4, containing 0.5% or 0.05% NP40 as stated in text     

Polymorphism in V k 10 genes encoding L chains of antibodies bearing the Ars-A and A48 cross-reactive idiotypes

p-azophenylarsonate-specific antibodies of A/J mice which bear the Ars-A cross-reactive idiotype utilize the V _K–Ars–A gene segment, a member of the V _K 10 family. Southern hybridization of genomic DNA from several inbred strains using a probe from the 5 flanking region of the V _K–Ars–A gene demonstrated three patterns of restrictio fragment length polymorphisms (RFLP). Six genes corresponding to hybridizing bands were obtained from DNA libraries of C.AKR, PERU and A/J mice, and nucleotide sequence comparisons revealed two allelic groups: AKRI (Igk-V10.1 ^a ), AJ1 (Igk-V10.1 ^b ) and PERU1 (Igk-V10.1 ^c ); AKR2 (Igk-V10.2 ^a ), AJ2 (Igk-V10.2 ^b ), and PERU2 (Igk-V10.2 ^c ).The Igk-V10.1 ^b gene of the A/J strain is the V _k–Ars–A gene used in Ars-A idiotype-positive antibodies. The product of the C.AKR allele (Igk-V10.1 ^a ) contained four amino acid substitutions in CDR3 as compared with Igk-V10.1 ^b . These substitutions probably explain the failure of AKR mice and other strains with the same V_K10 RFLP pattern to provide in genetic crosses a L chain which, together with the A/J V _H–ArsA gene product, form Ars-A idiotype-positive antibodies. Also, the nucleotide sequence identity between the Igk-V10.1 ^c and Igk-V10.1 ^b alleles and the Igk-V10.2 ^c and Igk-V10.2 ^b alleles is significantly greater than that seen in comparisons with the Igk-V10.1 ^a and Igk-V10.2 ^a alleles, respectively, suggesting an evolutionary pathway similar to that of the linked Igk-J locus.BALB/c antibodies bearing the A48 regulatory idiotype contain L chains encoded by the BALB/c Igk-V10.1 ^b and Igk-V10.2 ^b alleles. Strongly A48 idiotype-positive antibodies utilize the Igk-V10.1 ^b chain, and weakly A48-positive antibodies use the Igk-V10.2 ^b L chain. The possible effects of amino acid substitutions specified by the Igk-V10.1 ^a , Igk-V10.1 ^c , Igk-V10.2 ^a , and Igk-V10.2 ^c alleles on their ability to provide L chains used in A48 idiotype-positive are discussed.The locus name, Igk-V28 (D'Hoostelaere et al. 1988), will be used in this report in place of the name, Igk-VSer, used in the original publications (Goldrick et al. 1985; Boyd et al. 1986; Gottlieb et al. 1986; Ponath et al. 1988). The four alleles described at the Igk-VSer locus (Igk-VSer ^a , Igk-VSer ^b , Igk-VSer ^c , and Igk-VSer ^d ) are referred to as Igk-V28 ^a , Igk-V28 ^b , Igk-V28 ^c , and Igk-V28 ^d , respectively.The nucleotide sequence data reported in this paper have been submitted to GenBank nucleotide sequence database and have been assigned the accession numbers M54903, M54904, M54905, M54906, M54907, and M54908. Address correspondence and offprint requests to : P. D. Gottlieb.     

Control of the antibody response of C57BL/6 Lyt-congenic strains to the Lyt-3.1 alloantigen by a gene linked to theLyt-2 locus

Congenic anti-Lyt-3.1 sera have recently been produced by immunizing B6-Lyt-2^a mice with thymocytes from either B6-Lyt-2^a, Lyt-3^a or B6-Lyt-2^a, Lyt-3^a, H-2^k mice (Boos et al. 1978). Surprisingly, mice of the congenic strain B6 failed to produce either anti-Lyt-2.1 or anti-Lyt-3.1 cytotoxic antibodies after identical immunizations. To determine the genetic basis for the difference in response to Lyt-3.1, (B6 × B6-Lyt-2^a)F_a mice and progeny of the backcross, (B6 × B6-Lyt-2^a)F₁ × B6-Lyt-2^a, were immunized with B6-Lyt-2^a, Lyt-3^a, H-2^k thymocytes. In addition, thymic biopsies of backcross progeny were performed and thymocytes tested for the Lyt-2.2 antigenic specificity. Results indicate that gene(s) governing the immune response to Lyt-3.1 is (are) linked to theLyt-2 locus, and that the responder allele (linked toLyt-2 ^a ) shows very poor penetrance in Lyt-2^a/Lyt-2^b mice.     

Immunoglobulin constant kappa gene alleles in twelve strains of mice

Genomic DNA for the immunoglobulin (Ig) constant kappa Igk-C gene region was amplified by the polymerase chain reaction (PCR) method and sequenced from twelve commonly used inbred mouse strains. PCR products were used directly as templates in dideoxy-DNA-sequencing, a method which avoids the sequencing errors caused by Taq polymerase, since no cloning step is required. In restriction fragment length polymorphism (RFLP) studies the SJL mouse strain has been shown to belong to a Igk-C allogroup different from other common inbred mouse strains. The BALB/c Igk-C region was sequenced earlier, but our Igk-C sequences clarify the situation and confirm the existence of three Igk-C alleles in inbred mice (Mus musculus domesticus). Mice belonging to the kappa (Igk) haplotype e (SJL) have allele c of the Igk-C gene. The strains belonging to the kappa haplotype [a albino strain, K subline (AKR), PL and d (C58)] have allele a, and all other eight strains belonging to three different Igk haplotypes (b, c, and f) use allele b of the gene. Allele b has at least one (possibly two) nucleotide differences from allele a in the Igk-C region, but five compared to allele c. The allelic sequences also predict two allotypic kappa polypeptide chains among twelve inbred strains. Alleles a and b encode identical polypetides, but allele c (SJL) has a conserved lysine to arginine substitution in residue 142.The nucleotide sequence data reported in this paper have been submitted to the EMBL nucleotide sequence database and have been assigned the accession numbers X67002-X67012.     

Mouse Fcγ RI: identification and functional characterization of five new alleles

 The mouse Fcgr1 gene encoding the high-affinity IgG receptor (FcγRI) exists as two known alleles, FcγRI-BALB and FcγRI-NOD, and these alleles exhibit functional differences. To determine whether other alleles exist in mouse strains, Fcgr1 coding regions from 35 strains of mice were sequenced and a further five alleles were identified. The FcγRI-BALB and NOD alleles are now designated the "a" and "d" alleles, respectively. Analysis of the five new alleles revealed that although no polymorphisms were observed in the two leader exons, nucleotide and subsequent amino acid changes were observed in the exons encoding the extracellular domains, and transmembrane and cytoplasmic tail. The cDNA of the seven alleles (a–g) were isolated and transiently transfected into COS cells, and IgG-binding studies were performed. Receptors encoded by four of the five new alleles (b, c, f, g) bound IgG2a with high affinity, displaying IgG binding characteristics similar to the a allele (previously FcγRI-BALB). The d allele (previously FcγRI-NOD) and the e allele [derived from Mus spretus (SPRET/Ei)] encoded receptors which showed broader specificity by binding monomeric IgG2a, IgG2b, and IgG3. Received: 26 May 1999 / Revised: 25 October 1999     

The organization of immunoglobulin variable kappa chain genes on mouse chromosome 6

One mouse with a known recombination (NAK) at the Igk locus on chromosome 6 and two new recombinants [B6.PL (7 NS) and B6.PL (85NS)] were examined using a series of probes, each of which is specific for a set of immunoglobulin (Ig) Vk genes. Under high stringency conditions, each probe detects from 1 to 19 Bam HI restriction endonuclease fragments (REFs) in genomic DNA by Southern transfer hybridization techniques. Analysis of the REF patterns indicate that the NAK recombination event occurred within the variable region of Igk. The REF patterns of the two B6.PL congenic mice provided two additional recombination events which could be examined. Although some of the REFs had shared mobility among the parental strains, at least 1 and up to 13 polymorphic REFs were present for a given probe among the NZB and AKR parental strains. The results from the NAK mouse indicate that at least some members of Vk4, Vk8, Vk10, and Vk21 were on one side of the recombination event linked to the Lyt-2 ^a and Igk-Efl ^a alleles of AKR, while the Vk9, Vk11, and Vk24 REF patterns came from the NZB parental strain linked to the Igk-Ef2 ^b (Vk1) allele. The two B6.PL congenics produced a refined map on the Lyt-2, Lyt-3 side of the Vk region. The B6.PL (85NS) mice retained the Vk21 REF pattern of the Lyt-2 ^a, Lyt-3 ^a donor strain PL/J, while displaying the C57BL/6 REF pattern for the other Vk gene groups tested. The B6.PL (75NS) mice retained the REF patterns of PL/J for Vk21 and Ef-1, indicating a third recombination. This indicates the Vk gene order is (Lyt-2; Vk21); Ef-1; (Vk4; Vk8; Vk10); and (Vk9; Vk11; Vk24; Ef-2).     

Structural and evolutionary comparisons of four alleles of the mouse Igk-J locus which encodes immunoglobulin kappa light chain joining (J K ) segments

The Igk-J locus of the mouse encodes the immunoglobulin light chain joining (J) segments. Four Igk-J alleles have been described on the basis of restriction enzyme length polymorphisms. The nucleotide sequences of the Igk-J ^a allele (type strain, C.C58), Igk-J ^c allele (type strain, SJL/J), and Igk-J ^d allele (type strain, SK/CamRk) have been determined and are compared with the previously reported Igk-J ^b allele sequence (type strain, BALB/c). The mouse sequences are also compared with published sequences for rat and human J _k sequences. Far more differences were found between the Igk-J ^a allele and the other mouse alleles than between any two of the latter. These result in two amino acid substitutions which distinguish the J2 and J3 ¹ segments of the Igk-J ^a allele from the other three alleles. Use of the Phylogenetic Analysis Using Parsimony program to generate a phylogenetic tree strongly indicates that after divergence from the rat ancestor, there appears to have been an early split between the Igk-J ^a allele and the evolutionary precursor of the other mouse alleles. There also appears to have been far less divergence from the ancestral condition in the Igk-J ^a allele than in the other alleles. Also, the presence of only one convergent mutation among the four mouse alleles provides strong evidence against any crossing over within the Igk-J locus during the history of these alleles. Finally, the differences in rates of evolution of the Igk-J alleles are in marked contrast to the relatively uniform rates of divergence of four alleles of a mouse V _k gene, Igk-VSer.     

Nucleotide sequence of a mouse Tcp-1 pseudogene: A nucleotide record for a t complex gene carried by an ancestor of the mouse

We have isolated clones of a processed pseudogene of mouse t complex polypeptide 1 (Tcp-1) and determined the nucleotide sequence of the pseudogene. The pseudogene was 1363 bp long and had no intron. The Tcp-1 pseudogene had 88.4% or 88.3% nucleotide identity to the mouse Tcp-1 cDNA of wild-type (Tcp-1)^bor t haplotype (Tcp-1)^a, and 87.5% identity to the rat Tcp-1 cDNA. On 12 nucleotide positions where the open reading frames (ORFs) of mouse Tcp-1 ^band Tcp-1 ^acDNAs have bp substitutions, the Tcp-1 pseudogene had 6 bp identical to Tcp-1 ^b, 5 bp identical to Tcp-1 ^aand 1 bp not identical to neither. On ten amino acid positions where TCP-1B and TCP-1A polypeptides have substitutions, deduced amino acids of the Tcp-1 pseudogene had four amino acids identical to TCP-1B, five amino acids identical to TCP-1A and one amino acid identical to neither. These results suggest that the ancestral mouse Tcp-1 gene would have had no significant difference between the resemblance to Tcp-1 ^band that to Tcp-1 ^abefore they were diverged and that amino acids of TCP-1B and TCP-1A would have been substituted in similar high rates.The nucleotide sequence data reported in this paper have been submitted to GenBank and have been assigned the accession number D00851.     

cDNA sequence of four purine nucleoside phosphorylase (Np) alleles in the mouse

The molecular basis of four electrophoretic and activity variants of purine nucleoside phosphorylase in the mouse was examined by amplification and sequence analysis of cDNA. Compared with the cDNA coding sequence for C3H/HeHa designated Np ^a , there were five nucleotide changes for C57BL/6J, Np ^b ; three for MOLF/Ei, Np ^c ; and five for SPRET-1, Np ^d . There was only a single codon change between Np ^a and Np ^b , the deduced substitution of threonine 176 by serine. Similarly, there was only a single codon change between Np ^a and Np ^c , resulting in substitution of methionine 258 by lysine. There were three codon changes between Np ^a and Np ^d , resulting in substitution of glutamate 22 by lysine, threonine 39 by alanine, and aspartate 152 by glutamate. These amino acid substitutions-neutral to neutral, neutral to basic, and acidic to basic—are in agreement with the electrophoretic properties of the gene products of Np ^a relative to Np ^b , Np ^c , and Np ^d previously described by isoelectric focusing. Codon differences were confirmed by PCRRFLP or single nucleotide primer extension analysis and extended to include the assignment of other strains as Np ^a : C3H/HeHa, DBA/2J, CLA, Posch-2; or Np ^b : C57BL/6J, C57L/J, C58/J. Both RFLP analysis of amplified genomic DNA and Southern analysis are consistent with single but unique Np alleles present in the C3H/HeHa and C57BL/6J genomes. As these data do not support the previous two-loci, Np-1 and Np-2, classification, we propose and employ a new single locus multiple allele classification for Np on the basis of the sequence analysis.     

Structure, sequence, and polymorphism of the Lyt-2 T cell differentiation antigen gene

We have determined the nucleotide sequence and genomic organization of the mouse Lyt-2 T lymphocyte differentiation antigen gene. This gene consists of five exons and four introns, and the organization roughly parallels the protein domains. Alternative splicing to include or exclude exon IV (encoding part of the cytoplasmic tail) results in two forms of mRNA and accounts for the difference in size between the alpha- and alpha'-chains of Lyt-2. The gene structure provides further evidence for the evolutionary relationship between Lyt-2 and immunoglobulin genes. Comparison of the nucleotide sequence of the Lyt-2.1 and Lyt-2.2 alleles shows a high degree of conservation, but indicates that a single nucleotide change and consequent amino acid substitution in the variable region-like domain accounts for the serologic difference between these two alleles.     

H-2-linked genes determine the level of the primary in vitro anti-Mls response

The level of cell proliferation and interleukin-2 (IL-2) production observed in an anti-Mls mixed lymphocyte reaction between spleen cells from H-2 compatible, Mls incompatible mouse strains is determined by the H-2 haplotype of the mouse combination. Thus, while AKR (H-2 ^k) spleen cells stimulated strong M1s^a responses in H-2^k responder cells, AKR H-2b spleen cells stimulated no or negligible M1s^a responses in responder cells from H-2 ^bmouse strains. This effect was observed at the levels of IL-2 production and cell proliferation. The magnitude of the response observed using F₁ (H-2 ^k/H-2 ^b) responder cells was found to be a function of stimulator rather than responder cells. The poor stimulatory capacity of AKRH-2 ^bspleen cells was also shown not to be due to the loss of the stimulatory Mls ^aallele during the construction of the congenic strain from AKR and C57BL/6 parental strains. Using stimulator cells from a second series of congenic mice, we found H-2 ^b(strain DLLP) again to represent a poorly Mls^a stimulatory H-2 haplotype. In addition, H-2^q (DBA/1) cells displayed very poor Mls^a stimulatory potential while H-2^d (D1.C) cells were efficient Mls^a stimulators. Again the effect was shown to be at the level of the stimulator cells. In toto, our findings indicate that the H-2 ^kand H-2 ^dhaplotypes encode strong Mls^a stimulatory potential while the H-2 ^band H-2 ^qhaplotypes determine poor Mls^a stimulatory potential in primary in vitro responses, measured as cell proliferation and IL-2 production.Abbreviations used in this paper: CTL cytotoxic T lymphocyte - IL-1 interleukin-1 - IL-2 interleukin-2 - MLR mixed lymphocyte reaction - NMS normal mouse serum     

Characterization and DNA sequence of the b6w2 allotype of the rabbit immunoglobulin kappa 1 light chain (b locus)

The b6w2 allotype of the constant region of the rabbit immunoglobulin kappa 1 (k1) light chain (b locus) was discovered in wild populations from northern Spain. At the serological level, the b6w2 allotype is characterized by the presentation of all b6-specific epitopes, while an allotypic determinant which is shared between the nominal b5 and b6 allotypes is lacking. The DNA fragment encoding the b6w2 allotype was amplified by means of the polymerase chain reaction, and sequenced directly by dideoxy-DNA-sequencing. When compared with the sequence of the nominal b6 allele, the b6w2 sequence differs at eleven nucleotide positions (96.5% similarity). This variation corresponds to amino acid replacements at 1) the three positions C-terminal to the peptidyl junction with the variable region (amino acid positions 109–111);2) the four positions N-terminal to the interdomain disulfide bond (167–170); and 3) two positions in the vicinity of the interchain disulfide bond (190 and 210). The nature and distribution of the observed nucleotide substitutions strongly suggest a possible role of the extra interdomain disulfide bond in the unusual evolutionary dynamics of the rabbit K1 light chain.The nucleotide sequence data reported in this paper have been submitted to the EMBL nucleotide sequence database and have been assigned the accession number Z48308     

Sequence characterization of alleles Gpi1-s a and Gpi1-s b at the glucose phosphate isomerase structural locus

The sequences of alleles Gpil-s ^a and Gpi1-s ^b at the glucose phosphate isomerase structural locus have been determined from cDNA of the mouse inbred strains 101/H Gpi1-s ^a and C3H/HeH Gpi1-s ^b by TR PCR and direct sequencing of the amplified products. Four individual nucleotide differences were observed between the two alleles. The difference at amino acid residue 94, (Gpi1-s ^a GAT Asp, Gpi1-s ^b AAT Asn) may account for the differing electrophoretic migration, isoelectric point, and thermostability of the two alleles. Two of the other observed differences in the coding region (amino acid residue 12 Leu, Gpi1-s ^a CTC, Gpi1-s ^b CTG and amino acid residue 17 Arg, Gpi1-s ^a CGC, Gpi1-s ^b CGT) are silent and do not affect the predicted amino acid residues on translation. The fourth observed difference is located within the 3 noncoding sequences of the cDNA. The change at amino acid residue 94 is associated with the presence of a Hinf1 restriction site in Gpi1-s ^b, which is absent in Gpi1-s ^a, and may be a useful method for determining this marker.     

Sequence analysis and HLA-DR genotyping of a novel HLA-DRw14 allele

Analysis of a Japanese population by oligonucleotide genotyping revealed that one Japanese HLA-DRw14 allele had a DRB1 genotype different from that of the known HLA-DRw14-related alleles, DRB1 ^* 1401 (DRw14-Dw9) and DRB1 ^* 1402 (DRw14-Dw16). The second exon of the DRB1 gene of the novel DRw14 allele (designated DRB1-14c) was amplified enzymatically and sequenced after cloning intto a plasmid vector. The amino acid sequence of the first domain in the DR1 chain encoded in the DRB1-14c allele was more similar to that of the DRB1 ^* 1401 allele (three amino acid substitutions).than to that of the DRB1 ^* 1402 allele (six amino acid substitutions). No polymorphic amino acid residue that could explain the common serologic HLA-DRw14 specificity was identified among the sequences of the three DRw14-related alleles. Sequence-specific oligonucleotides (SSOs) were synthesized on the basis of the DRB1-14c nucleotide sequence and used for genotyping of the Japanese population. These SSOs served as useful probes for identifying the DRB1-14c allele in a wide range of donors.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number M33693.     

Isolation and Analysis of Rice Rf1-Orthologus PPR Genes Co-segregating with <Emphasis Type="Italic">Rf3</Emphasis> in Maize

Using an in silico cloning approach, five putative maize pentatricopeptide repeat (PPR)-containing protein genes (PPR-814a, PPR-814b, PPR-814c, PPR-816, PPR-817) with complete open reading frames were identified in the inbred line S-Mo17^Rf3Rf3. The amino acid sequence indicated that these genes encoded mitochondrially targeted proteins containing repeats of a 35-aa PPR motif. The genes were mapped into the interval umc1525–bnlg1520 on chromosome 2. In a non-restoring genotype, we identified three homologous genes that contained deletions or nucleotide substitutions in the coding region. Sequence analysis revealed that one of the three genes (PPR-814a, PPR-814b, PPR-814c) could be considered a candidate restorer gene for S male sterility cytoplasm, and linkage analysis demonstrated that the genes co-segregated with the fertility restorer gene Rf3.     

Polymorphisms in the coding and noncoding regions of murinePgk-1 alleles

The mouse X-linkedPgk-1 gene encodes phosphoglycerate kinase. When transfected into human cells, thePgk-1^b allele causes the appearance of mouse PGK-1^b enzyme activity. We describe here cloning of mousePgk-1^a, an allele ofPgk-1 which encodes an enzyme, PGK-1^a, with distinct electrophoretic mobility. We constructed recombinants between the DNA encodingPgk-1^b andPgk-1^a and transfected these constructs into human to assess the electrophoretic characteristics of each recombinant. In this way the charge variation between the two proteins was localized to exons 4 or 5. Sequencing of these exons revealed a single base-pair difference between the two alleles at codon 155, which predicts the amino acids lysine and threonine in PGK-1^b and PGK-1^a, respectively. A number of other DNA sequence polymorphisms exist betweenPgk-1^b andPgk-1^a including part of an L1 repeated element unique toPgk-1^a. This work was supported by the Medical Research Council of Canada, the National Cancer Institute of Canada, and the Deutsche Forschungsgemeinschaft, SFB 304.     

Polymorphism and Expression of cDNAs Encoding Glycinin Subunits

The nucleotide sequence of cDNA encoding the glycinin A₂B_1a subunit from var. Shirotsurunoko was determined and compared with that in the case of var. Bonminori. The comparison showed six nucleotide substitutions in the coding sequence, one of which results in one amino acid replacement, and three in the 3'-noncoding region. These differences indicate the occurrence of polymorphism of the glycinin A₂B_1a subunit gene between the cultivars. The present data together with the previous results indicating the polymorphism of the A_1aB_1b subunit gene [(Utsumi et al., J. Agric. Food Chem., 35, 210 (1987)] suggest that the polymorphism is a general property of glycinin subunit genes. The expression of cDNAs encoding the A₂B_1a and A_1aB_1b subunits was examined. The results obtained in both in vivo- and in vitro-expression experiments indicate that the resultant products were readily degraded.