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Table 1.

Sequences of degenerate primers used for cloning of short gastrulation (sog), single-minded (sim), prospero (pros), engrailed (en) and optomotor blind (omb) genes

Primer nameNucleotide sequence(5′-3′)*Amino acid sequence
sog1 tggcayccnttygtnccncc WHPFVPP 
sog2 ggrcaytcyttrcarcankc (A/D)CCKQCP 
sog3 gayytnggnccnccnttyg DLGPPFG 
sog4 acncknckccanacnccrca CGVWRRV 
sog5 tgymrnaayathaaraaygantg C(K/R)NIKN(D/E)C 
sog6 ccnggrcangtyttrcarca CCKTCPG 
sim1 gnatgaartgygtnytngc RMKCVLAK 
sim2 tanswytgnacccanaccca WVWVQSY 
sim3 ccnswrcartgdatnacytt KVIHCSG 
pros1 aargcnaarytnatgttyttyt KAKLMFF(W/Y) 
pros2 gcytgnckngcrwayttytc EK(Y/F)ARQA 
en1 tggccngcntgggtntwytg WPAWV(F/Y)C 
en2 ttrtanarnccytgngccat MAQGLYN 
en3 aakgakaatmgktatttkackga (N/Q)(D/E)NRYLTE 
en4 ttrttytgraaccadatytt KIWFQNK 
omb1 ayggnmgnmgnatgttycc NGRRMFP 
omb2 aangcyttngcraanggrtt NPFAKAF 
Primer nameNucleotide sequence(5′-3′)*Amino acid sequence
sog1 tggcayccnttygtnccncc WHPFVPP 
sog2 ggrcaytcyttrcarcankc (A/D)CCKQCP 
sog3 gayytnggnccnccnttyg DLGPPFG 
sog4 acncknckccanacnccrca CGVWRRV 
sog5 tgymrnaayathaaraaygantg C(K/R)NIKN(D/E)C 
sog6 ccnggrcangtyttrcarca CCKTCPG 
sim1 gnatgaartgygtnytngc RMKCVLAK 
sim2 tanswytgnacccanaccca WVWVQSY 
sim3 ccnswrcartgdatnacytt KVIHCSG 
pros1 aargcnaarytnatgttyttyt KAKLMFF(W/Y) 
pros2 gcytgnckngcrwayttytc EK(Y/F)ARQA 
en1 tggccngcntgggtntwytg WPAWV(F/Y)C 
en2 ttrtanarnccytgngccat MAQGLYN 
en3 aakgakaatmgktatttkackga (N/Q)(D/E)NRYLTE 
en4 ttrttytgraaccadatytt KIWFQNK 
omb1 ayggnmgnmgnatgttycc NGRRMFP 
omb2 aangcyttngcraanggrtt NPFAKAF 

Achaearanea sog (At-sog) was amplified using the sog1 and sog2 primers, and Pholcus sog (Pp-sog) and Artemia sog (Af-sog) were amplified using the sog3 and sog4 primers for first PCR and the sog5 and sog6 primers for nested PCR. Achaearanea sim (At-sim) was amplified using the sim1 and sim2 primers for first PCR and the sim1 and sim3 primers for nested PCR. Achaearanea pros (At-pros) was amplified using the pros1 and pros2 primers. Achaearanea en (At-en) was amplified using the en1 and en2 primers for first PCR and the en3 and en4 primers for nested PCR. Achaearanea omb (At-omb) was amplified using the omb1 and omb2 primers. The PCR conditions were as follows: 1 cycle of 95°C for 5 minutes, 50°C for 2 minutes 30 seconds, 72°C for 40 seconds; 35 cycles of 95°C for 40 seconds, 50°C for 40 seconds, 72°C for 40 seconds;1 cycle of 72°C for 10 minutes; and then a 4°C soak. For amplification of At-en and At-omb, the annealing temperatures were 45°C and 40°C, respectively, instead of 50°C. To obtain full-length cDNAs for At-sog At-en, At-omb and Artemia sog,cDNA libraries (Akiyama-Oda and Oda,2003; Oda et al.,2005) were screened with DIG-labeled DNA probes for the PCR-amplified fragments. To obtain full-length cDNAs for At-sim,At-pros and Pp-sog, 5′ and 3′ RACE were performed using a SMART RACE cDNA amplification kit (Clontech) and ExTaq polymerase(Takara)

*

d is a, g or t; h is a, c or t; k is g or t; m is a or c; n is a, c, g or t; r is a or g; s is c or g; w is a or t; y is t or c

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