HindIII R6041

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	The clone 4G4E4.0 encompassing a 4-kb EcoRI 59 fragment of the human huntingtin gene (including exon 1 and upstream noncoding region) (Lin et al., 1995) was characterized by restriction endonuclease mapping with EcoRI, HindIII, and SfiI. A region of 3345 nt of the 59-untranslated region was deleted by double digestion with HindIII and SfiI to form a human huntingtin exon 1 transcription plasmid (clone 839; Fig. 1).

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The clone 4G4E4.0 encompassing a 4-kb EcoRI 59 fragment of the human huntingtin gene (including exon 1 and upstream noncoding region) (Lin et al., 1995) was characterized by restriction endonuclease mapping with EcoRI, HindIII, and SfiI. A region of 3345 nt of the 59-untranslated region was deleted by double digestion with HindIII and SfiI to form a human huntingtin exon 1 transcription plasmid (clone 839; Fig. 1).

Publication protocol

"The construction of an expression vector
containing the human huntingtin exon 1 with a normal range repeat
for in vitro transcription is summarized in Fig. 1. The clone 4G4E4.0
encompassing a 4-kb EcoRI 59 fragment of the human huntingtin
gene (including exon 1 and upstream noncoding region) (Lin et al.,
1995) was characterized by restriction endonuclease mapping with
EcoRI, HindIII, and SfiI. A region of 3345 nt of the 59-untranslated
region was deleted by double digestion with HindIII and SfiI to form
a human huntingtin exon 1 transcription plasmid (clone 839; Fig. 1).
An mRNA containing 222 nt of the 59-untranslated region of human
huntingtin exon 1 and an open reading frame that translates into a
10,068-Da huntingtin gene product corresponding to human exon 1
was used for in vitro expression driven by a T3 promoter (Fig. 1). The
Qiagen maxi-prep plasmid isolation system (Santa Clara, CA) was
used for plasmid preparation."

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