Table 1.

Description of the Probes for Each Globin Type Used in the RNase Protection Assays

Globin Type Transcription Vector RE/PromoterProbe Length (bases) Protected Fragment (bases)Region Protected
βmaj/min βS/T pCR2.1  HindIII/T7  241  113 Part of exon I Part of exon II  
ε  pBluescript KS XbaI/T3  348  253  Part of exon II Entire exon III Part of 3′UTR  
βh1  pSP65 HindIII/Sp6  379  245 or 180  Part of exon III Part of 3′UTR  
α-1/α-2  pBluescript SK XbaI/T7  193  96  Part of exon II  
ζ pSP64  EcoRI/Sp6  239  157  Part of exon I 
Globin Type Transcription Vector RE/PromoterProbe Length (bases) Protected Fragment (bases)Region Protected
βmaj/min βS/T pCR2.1  HindIII/T7  241  113 Part of exon I Part of exon II  
ε  pBluescript KS XbaI/T3  348  253  Part of exon II Entire exon III Part of 3′UTR  
βh1  pSP65 HindIII/Sp6  379  245 or 180  Part of exon III Part of 3′UTR  
α-1/α-2  pBluescript SK XbaI/T7  193  96  Part of exon II  
ζ pSP64  EcoRI/Sp6  239  157  Part of exon I 

After each transcription vector containing the globin DNA fragment was digested with the RE, an antisense RNA probe was transcribed with the specified promoter. The length (base number) of the probe and the protected fragment, and the region protected are summarized. The length of protected fragment of βh1-globin mRNA differed depending on the haplotype, consisting of 245 bases in the diffuse haplotype (Ba/F3 cells and 11.5-day-old DBA/2 embryonic blood) and 180 bases in the single haplotype (B6SUtA cells), as discussed in the text.

Abbreviation: RE, restriction endonuclease.