28
G. Jansen et al.
genes per cell. We can now address questions such as how specificity is
achieved and maintained in the presence of so many, probably rather
similar signal transduction cascades, and where the information is integrated.
2.3 Methods
2.3.1 Ga Gene Identification
The C. elegans genomic sequence, available from the Genome Sequencing Centers (The Sanger Centre, Cambridge, UK, and Washington University, St. Louis, Mo. USA), was repeatedly screened for Ga subunit
genes in a TBLASTN search (Altschul et al. 1990).The DNA sequences
were further analyzed using the Wisconsin Sequence Analysis Package.
The predicted gene structures were further refined using the GENEFINDER predictions, as annotated in the C. elegans database ACeDB
(Eeckman and Durbin 1995). Discrepancies between the GENEFINDER predictions and ours were communicated to those annotating the
sequences.
We identified 20 G protein a subunit genes in the C. elegans genome
(Jansen et al. 1999): gpa-l (Lochrie et al. 1991), located on cosmid
T19C4, gene 6 (i.e., T19C4.6); gpa-2 (Zwaal et al. 1997), F38E1.5;
gpa-3 (Zwaal et al. 1997), E02CI2.5; gpa-4, T07A9.m; gpa-5,
F53B1.7; gpa-6, F48Cl1.1; gpa-7, RlOHlO.5; gpa-8, F56H9.3; gpa-9,
F56H9.4; gpa-lO, C55H1.2; gpa-ll, C16A11.1; gpa-12, F18G5.3; gpa13, CI5C8.7; gpa-l4, B0207.3; gpa-l5, M04C7.1; gpa-l6, Y95B8;
odr-3 (Roayaie et al. 1998), C34D1.3; goa-l (Lochrie et al. 1991),
C26C6.2; gsa-1 (Park et al. 1997), R06AlO.2; egl-30 (Brundage et al.
1996), MOl D7. 7. Furthermore, two G~ and two Gy subunits were
identified: gpb-l (Zwaal et al. 1996), F13Dl2.7; gpb-2, F52A8.2; gpc-l,
K02A4.2; gpc-2, F08B6.e. None of these genes seems part of an operon.
The gene structure prediction of gpa-5 and -16 was confirmed by the
sequencing of random and oligo-dT primed cDNA amplified by reverse
transcriptase PCR.
G. Jansen et al.
genes per cell. We can now address questions such as how specificity is
achieved and maintained in the presence of so many, probably rather
similar signal transduction cascades, and where the information is integrated.
2.3 Methods
2.3.1 Ga Gene Identification
The C. elegans genomic sequence, available from the Genome Sequencing Centers (The Sanger Centre, Cambridge, UK, and Washington University, St. Louis, Mo. USA), was repeatedly screened for Ga subunit
genes in a TBLASTN search (Altschul et al. 1990).The DNA sequences
were further analyzed using the Wisconsin Sequence Analysis Package.
The predicted gene structures were further refined using the GENEFINDER predictions, as annotated in the C. elegans database ACeDB
(Eeckman and Durbin 1995). Discrepancies between the GENEFINDER predictions and ours were communicated to those annotating the
sequences.
We identified 20 G protein a subunit genes in the C. elegans genome
(Jansen et al. 1999): gpa-l (Lochrie et al. 1991), located on cosmid
T19C4, gene 6 (i.e., T19C4.6); gpa-2 (Zwaal et al. 1997), F38E1.5;
gpa-3 (Zwaal et al. 1997), E02CI2.5; gpa-4, T07A9.m; gpa-5,
F53B1.7; gpa-6, F48Cl1.1; gpa-7, RlOHlO.5; gpa-8, F56H9.3; gpa-9,
F56H9.4; gpa-lO, C55H1.2; gpa-ll, C16A11.1; gpa-12, F18G5.3; gpa13, CI5C8.7; gpa-l4, B0207.3; gpa-l5, M04C7.1; gpa-l6, Y95B8;
odr-3 (Roayaie et al. 1998), C34D1.3; goa-l (Lochrie et al. 1991),
C26C6.2; gsa-1 (Park et al. 1997), R06AlO.2; egl-30 (Brundage et al.
1996), MOl D7. 7. Furthermore, two G~ and two Gy subunits were
identified: gpb-l (Zwaal et al. 1996), F13Dl2.7; gpb-2, F52A8.2; gpc-l,
K02A4.2; gpc-2, F08B6.e. None of these genes seems part of an operon.
The gene structure prediction of gpa-5 and -16 was confirmed by the
sequencing of random and oligo-dT primed cDNA amplified by reverse
transcriptase PCR.