Description of the Probes for Each Globin Type Used in the RNase Protection Assays
Globin Type . | Transcription Vector . | RE/Promoter . | Probe 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/Promoter . | Probe 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.